thioredoxin reductase  (Millipore)


Bioz Verified Symbol Millipore is a verified supplier  
  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 94
    Name:
    NADPH
    Description:
    Amorphous powder bottled under nitrogen
    Catalog Number:
    nadph-ro
    Price:
    None
    Applications:
    NADPH-RO has been used for the determination of thioredoxin reductase activity.
    Buy from Supplier


    Structured Review

    Millipore thioredoxin reductase
    NADPH
    Amorphous powder bottled under nitrogen
    https://www.bioz.com/result/thioredoxin reductase/product/Millipore
    Average 94 stars, based on 1188 article reviews
    Price from $9.99 to $1999.99
    thioredoxin reductase - by Bioz Stars, 2020-09
    94/100 stars

    Images

    1) Product Images from "Sodium selenite supplementation does not fully restore oxidative stress-induced deiodinase dysfunction: Implications for the nonthyroidal illness syndrome"

    Article Title: Sodium selenite supplementation does not fully restore oxidative stress-induced deiodinase dysfunction: Implications for the nonthyroidal illness syndrome

    Journal: Redox Biology

    doi: 10.1016/j.redox.2015.09.002

    Effect of increasing levels of sodium selenite (0–300 nM) on glutathione peroxidase activity (A), thioredoxin reductase (TRx) activity (B) and GSH (C) levels in MSTO-211 (●), HepG2 (▲) or MCF-7 (■) cells. Data are mean±SD of at least three independent experiments.* P
    Figure Legend Snippet: Effect of increasing levels of sodium selenite (0–300 nM) on glutathione peroxidase activity (A), thioredoxin reductase (TRx) activity (B) and GSH (C) levels in MSTO-211 (●), HepG2 (▲) or MCF-7 (■) cells. Data are mean±SD of at least three independent experiments.* P

    Techniques Used: Activity Assay

    Effect of sodium selenite or N-acetylcysteine (NAC) on glutathione peroxidase (GPx) and thioredoxin reductase (TRx) activity in controls and IL6 (1000 ng/L) treated cells. The column on the left shows the effect of sodium selenite or NAC on intracellular GSH levels in controls and IL6 (1000 ng/L) treated MSTO-211 (A), HepG2 (B), or MCF-7 (C) cells. The column on the right shows the effect of sodium selenite or NAC on intracellular TRx levels in MSTO-211 (D), HepG2 (E), or MCF-7 (F) cells. Data are mean±SD of at least three independent experiments.
    Figure Legend Snippet: Effect of sodium selenite or N-acetylcysteine (NAC) on glutathione peroxidase (GPx) and thioredoxin reductase (TRx) activity in controls and IL6 (1000 ng/L) treated cells. The column on the left shows the effect of sodium selenite or NAC on intracellular GSH levels in controls and IL6 (1000 ng/L) treated MSTO-211 (A), HepG2 (B), or MCF-7 (C) cells. The column on the right shows the effect of sodium selenite or NAC on intracellular TRx levels in MSTO-211 (D), HepG2 (E), or MCF-7 (F) cells. Data are mean±SD of at least three independent experiments.

    Techniques Used: Activity Assay

    2) Product Images from "GlnR Activation Induces Peroxide Resistance in Mycobacterial Biofilms"

    Article Title: GlnR Activation Induces Peroxide Resistance in Mycobacterial Biofilms

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2018.01428

    Role of gpr in maintenance of redox homeostasis of M. smegmatis. (A,B) Levels of oxidized (NADP) form of nicotinamide adenine dinucleotide phosphate in biofilms of wild-type, Δ gpr and the complemented strains cultured in normal Sauton’s medium. (C) Ratio of NADPH to NADP calculated from (A,B) . Data represent mean of three biologically independent experiments ∗ , ∗∗ , and ∗∗∗ indicate p ( t -test)
    Figure Legend Snippet: Role of gpr in maintenance of redox homeostasis of M. smegmatis. (A,B) Levels of oxidized (NADP) form of nicotinamide adenine dinucleotide phosphate in biofilms of wild-type, Δ gpr and the complemented strains cultured in normal Sauton’s medium. (C) Ratio of NADPH to NADP calculated from (A,B) . Data represent mean of three biologically independent experiments ∗ , ∗∗ , and ∗∗∗ indicate p ( t -test)

    Techniques Used: Cell Culture

    3) Product Images from "Two enzymes with redundant fructose bisphosphatase activity sustain gluconeogenesis and virulence in Mycobacterium tuberculosis"

    Article Title: Two enzymes with redundant fructose bisphosphatase activity sustain gluconeogenesis and virulence in Mycobacterium tuberculosis

    Journal: Nature communications

    doi: 10.1038/ncomms8912

    Metabolic schematic of glycolysis and gluconeogenesis GlpX  ( rv1099c ) encodes the only known FBPase in the  Mtb  genome and controls the rate-limiting step of gluconeogenesis. Other enzymes of the gluconeogenesis pathway are encoded by  pckA  (phosphoenolpyruvate carboxykinase),  eno  (enolase),  gpm1  (phosphoglycerate mutase),  pgk  (phosphoglycerate kinase),  gap  (glyceraldehyde 3-phosphate dehydrogenase),  tpi  (triose phosphate isomerase),  fba  (fructose bisphosphate aldolase) and  pgi  (glucose 6-phosphate isomerase). Also shown are  ppgK  (polyphosphate glucokinase),  glkA  (glucokinase) and  pfkA  and  pfkB  (phosphofructokinases), which encode glycolysis-specific enzymes. G6P: glucose 6-phosphate, F6P: fructose 6-phosphate, FBP: fructose 1,6-bisphosphate, DHAP: dihydroxyacetone phosphate, G3P: glyceraldehyde 3-phosphate, 1,3BPG: 1,3-bisphosphoglycerate, 3PG: 3-phosphoglycerate, 2PG: 2-phosphoglycerate, PEP: phosphoenolpyruvate, PYR: pyruvate, OAA: oxaloacetate.
    Figure Legend Snippet: Metabolic schematic of glycolysis and gluconeogenesis GlpX ( rv1099c ) encodes the only known FBPase in the Mtb genome and controls the rate-limiting step of gluconeogenesis. Other enzymes of the gluconeogenesis pathway are encoded by pckA (phosphoenolpyruvate carboxykinase), eno (enolase), gpm1 (phosphoglycerate mutase), pgk (phosphoglycerate kinase), gap (glyceraldehyde 3-phosphate dehydrogenase), tpi (triose phosphate isomerase), fba (fructose bisphosphate aldolase) and pgi (glucose 6-phosphate isomerase). Also shown are ppgK (polyphosphate glucokinase), glkA (glucokinase) and pfkA and pfkB (phosphofructokinases), which encode glycolysis-specific enzymes. G6P: glucose 6-phosphate, F6P: fructose 6-phosphate, FBP: fructose 1,6-bisphosphate, DHAP: dihydroxyacetone phosphate, G3P: glyceraldehyde 3-phosphate, 1,3BPG: 1,3-bisphosphoglycerate, 3PG: 3-phosphoglycerate, 2PG: 2-phosphoglycerate, PEP: phosphoenolpyruvate, PYR: pyruvate, OAA: oxaloacetate.

    Techniques Used:

    4) Product Images from "Orchestrated Domain Movement in Catalysis by Cytochrome P450 Reductase"

    Article Title: Orchestrated Domain Movement in Catalysis by Cytochrome P450 Reductase

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-09840-8

    Analysis of SANS data in terms of multiple states. ( a – c ) Fits to the scattering curve for CPR reduced to the 2e − level with dithionite using one, two or three states (from a 10,000 conformation pool) respectively. ( d , e ) Fits to the scattering curve for CPR reduced to the 2e − level with dithionite using a two-state model; the extended state was represented by, ( d ) the model of Hamdane et al . 42 or, ( e ) the model of Huang et al . 37 ; in both cases the crystal structure was used as a model for the compact state. ( f , g ) Fits to the scattering curve for CPR reduced to the 2e − level with NADPH using a two-state model; the extended state was represented by ( f ) the model of Hamdane et al . 42 or ( g ) the model of Huang et al . 37 ; in both cases the crystal structure was used as a model for the compact state. In all cases the goodness-of-fit is indicated by the χ 2 value.
    Figure Legend Snippet: Analysis of SANS data in terms of multiple states. ( a – c ) Fits to the scattering curve for CPR reduced to the 2e − level with dithionite using one, two or three states (from a 10,000 conformation pool) respectively. ( d , e ) Fits to the scattering curve for CPR reduced to the 2e − level with dithionite using a two-state model; the extended state was represented by, ( d ) the model of Hamdane et al . 42 or, ( e ) the model of Huang et al . 37 ; in both cases the crystal structure was used as a model for the compact state. ( f , g ) Fits to the scattering curve for CPR reduced to the 2e − level with NADPH using a two-state model; the extended state was represented by ( f ) the model of Hamdane et al . 42 or ( g ) the model of Huang et al . 37 ; in both cases the crystal structure was used as a model for the compact state. In all cases the goodness-of-fit is indicated by the χ 2 value.

    Techniques Used:

    Effects of salt concentration on the kinetics of ET by CPR and on its conformation. ( a ) Stopped-flow traces showing the reduction of cytochrome c on rapid mixing of CPR pre-reduced with NADPH and cytochrome c , at various concentrations of added salt; the percentage of a single turnover which occurred within the 2ms deadtime of the instrument is plotted as a function of salt in ( b ). ( c ) Pairwise distance distribution functions derived from the SANS data at different salt concentrations, showing that increasing the salt concentration leads to a more extended conformation, in qualitative agreement with SAXS studies 37 , 45 . ( d ) Porod-Debye plots, showing increased flexibility of CPR at ≥0.6 M NaCl.
    Figure Legend Snippet: Effects of salt concentration on the kinetics of ET by CPR and on its conformation. ( a ) Stopped-flow traces showing the reduction of cytochrome c on rapid mixing of CPR pre-reduced with NADPH and cytochrome c , at various concentrations of added salt; the percentage of a single turnover which occurred within the 2ms deadtime of the instrument is plotted as a function of salt in ( b ). ( c ) Pairwise distance distribution functions derived from the SANS data at different salt concentrations, showing that increasing the salt concentration leads to a more extended conformation, in qualitative agreement with SAXS studies 37 , 45 . ( d ) Porod-Debye plots, showing increased flexibility of CPR at ≥0.6 M NaCl.

    Techniques Used: Concentration Assay, Flow Cytometry, Derivative Assay

    Structural information from SANS data on the different redox states of CPR. ( a , b ) Pairwise distance distribution functions; ( a) samples reduced with dithionite; ( b ) samples reduced with NADPH. The longer ‘tail’ on the distribution functions for the reduced states as compared to the oxidised state demonstrates a more extended average conformation. ( c , d ) ab initio envelopes calculated using DAMMIF from the scattering curves of ( c ) oxidised CPR and ( d ) CPR reduced to the 2e − level with dithionite; in ( c and d ) the lower envelopes have been rotated 90° about the horizontal axis.
    Figure Legend Snippet: Structural information from SANS data on the different redox states of CPR. ( a , b ) Pairwise distance distribution functions; ( a) samples reduced with dithionite; ( b ) samples reduced with NADPH. The longer ‘tail’ on the distribution functions for the reduced states as compared to the oxidised state demonstrates a more extended average conformation. ( c , d ) ab initio envelopes calculated using DAMMIF from the scattering curves of ( c ) oxidised CPR and ( d ) CPR reduced to the 2e − level with dithionite; in ( c and d ) the lower envelopes have been rotated 90° about the horizontal axis.

    Techniques Used:

    5) Product Images from "Methanococci Use the Diaminopimelate Aminotransferase (DapL) Pathway for Lysine Biosynthesis ▿"

    Article Title: Methanococci Use the Diaminopimelate Aminotransferase (DapL) Pathway for Lysine Biosynthesis ▿

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.00172-10

    DapL activity (A) and heat stability (B) of MJ1391. (A) The DapL assay was performed at 70°C for 0 to 60 min before termination and measurement of the product glutamate. (B) The temperature stabilities of MJ1391 and the substrates α-KG
    Figure Legend Snippet: DapL activity (A) and heat stability (B) of MJ1391. (A) The DapL assay was performed at 70°C for 0 to 60 min before termination and measurement of the product glutamate. (B) The temperature stabilities of MJ1391 and the substrates α-KG

    Techniques Used: Activity Assay

    6) Product Images from "Characterization of pH dependent Mn(II) oxidation strategies and formation of a bixbyite-like phase by Mesorhizobium australicum T-G1"

    Article Title: Characterization of pH dependent Mn(II) oxidation strategies and formation of a bixbyite-like phase by Mesorhizobium australicum T-G1

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2015.00734

    Characterization of biogenic Mn oxides produced by strain T-G1 in PYG medium at pH 5.5. (A) Scanning electron micrograph showing spherical clusters of micron to sub-micron sheets of Mn oxide minerals. (B) An enlargement of the area indicated by a red box in (A) . (C) Confocal laser scanning microscopy (CLSM) of a mineral aggregate in association with T-G1. Bacterial cells stained by FM 1-43 probe shown in green, reflection of minerals was indicated in gray. (D) Energy-dispersive X-ray (EDS) spectra showing elemental composition of biogenic Mn oxide particles. (E) Raman spectroscopy of biogenic Mn oxides. (F) XRD pattern of biogenic Mn-oxide minerals with vertical lines indicating strong (red) and weak peak positions (green) of bixbyite.
    Figure Legend Snippet: Characterization of biogenic Mn oxides produced by strain T-G1 in PYG medium at pH 5.5. (A) Scanning electron micrograph showing spherical clusters of micron to sub-micron sheets of Mn oxide minerals. (B) An enlargement of the area indicated by a red box in (A) . (C) Confocal laser scanning microscopy (CLSM) of a mineral aggregate in association with T-G1. Bacterial cells stained by FM 1-43 probe shown in green, reflection of minerals was indicated in gray. (D) Energy-dispersive X-ray (EDS) spectra showing elemental composition of biogenic Mn oxide particles. (E) Raman spectroscopy of biogenic Mn oxides. (F) XRD pattern of biogenic Mn-oxide minerals with vertical lines indicating strong (red) and weak peak positions (green) of bixbyite.

    Techniques Used: Produced, Confocal Laser Scanning Microscopy, Staining, Raman Spectroscopy

    Phylogenetic tree showing the evolutionary placement of Mesorhizobium australicum T-G1 (in rectangle) relative to known MOB based on 16S rRNA gene sequences . black squares indicate non-Mn(II)-oxidizing strains. The tree was built using a neighbor joining algorithm with bootstrap values, displayed as percentages, based on 1000 replicates. Accession numbers for published sequences are shown in parentheses. The scale bar represents 0.02 substitutions per nucleotide position.
    Figure Legend Snippet: Phylogenetic tree showing the evolutionary placement of Mesorhizobium australicum T-G1 (in rectangle) relative to known MOB based on 16S rRNA gene sequences . black squares indicate non-Mn(II)-oxidizing strains. The tree was built using a neighbor joining algorithm with bootstrap values, displayed as percentages, based on 1000 replicates. Accession numbers for published sequences are shown in parentheses. The scale bar represents 0.02 substitutions per nucleotide position.

    Techniques Used:

    Mn(II) oxidation and growth of T-G1 at different pH values. Growth (OD 600 ) and Mn(II) oxidation at pH 5.5 and pH 7.2. The data for the Mn(II) oxidation control at pH 7.2 overlapped with that at pH 5.5, therefore, only the data at pH 5.5 is shown. Error bars represent SD.
    Figure Legend Snippet: Mn(II) oxidation and growth of T-G1 at different pH values. Growth (OD 600 ) and Mn(II) oxidation at pH 5.5 and pH 7.2. The data for the Mn(II) oxidation control at pH 7.2 overlapped with that at pH 5.5, therefore, only the data at pH 5.5 is shown. Error bars represent SD.

    Techniques Used:

    Morphology and Mn(II) oxidation by strain T-G1: (A) scanning electron micrograph of T-G1 cells, showing abundant extracellular polymeric substances; (B) Mn(II) oxidation by T-G1 as seen by a color change in PYG medium with 10 mM MnSO 4 (right) and without Mn (left); and (C) leucoberbelin blue (LBB) spot test on colony of T-G1 showing a blue color change indicative of Mn oxide production .
    Figure Legend Snippet: Morphology and Mn(II) oxidation by strain T-G1: (A) scanning electron micrograph of T-G1 cells, showing abundant extracellular polymeric substances; (B) Mn(II) oxidation by T-G1 as seen by a color change in PYG medium with 10 mM MnSO 4 (right) and without Mn (left); and (C) leucoberbelin blue (LBB) spot test on colony of T-G1 showing a blue color change indicative of Mn oxide production .

    Techniques Used: Spot Test

    7) Product Images from "Mechanisms underlying the predictive power of high skeletal muscle uptake of FDG in amyotrophic lateral sclerosis"

    Article Title: Mechanisms underlying the predictive power of high skeletal muscle uptake of FDG in amyotrophic lateral sclerosis

    Journal: EJNMMI Research

    doi: 10.1186/s13550-020-00666-6

    Mitochondrial/ER ultrastructure and bioenergetics activity in skeletal muscles. a , b Electron microscopy representative images of control and SOD1 G93A quadriceps ( n = 3). In both panels, red arrows show mitochondria while yellow arrows show ER ultrastructural profiles. c – e Western blot analysis and relative densitometry quantitative analyses of Mfn2, Drp1, and Calnexin in the two groups. f , g ATP synthesis and oxygen consumption rate stimulated by pyruvate/malate in control (green column) and SOD1 G93A (red column) of isolated mitochondria. h Complex I activity in isolated mitochondria form control (green column) and SOD1 G93A muscles (red column) studied by the FeCN reduction in the presence of NADH. Data are expressed as mean ± SD, n = 3 for each group. Student t test for unpaired data was used for statistical evaluation
    Figure Legend Snippet: Mitochondrial/ER ultrastructure and bioenergetics activity in skeletal muscles. a , b Electron microscopy representative images of control and SOD1 G93A quadriceps ( n = 3). In both panels, red arrows show mitochondria while yellow arrows show ER ultrastructural profiles. c – e Western blot analysis and relative densitometry quantitative analyses of Mfn2, Drp1, and Calnexin in the two groups. f , g ATP synthesis and oxygen consumption rate stimulated by pyruvate/malate in control (green column) and SOD1 G93A (red column) of isolated mitochondria. h Complex I activity in isolated mitochondria form control (green column) and SOD1 G93A muscles (red column) studied by the FeCN reduction in the presence of NADH. Data are expressed as mean ± SD, n = 3 for each group. Student t test for unpaired data was used for statistical evaluation

    Techniques Used: Activity Assay, Electron Microscopy, Western Blot, Isolation

    Mitochondrial ultrastructure and bioenergetics activity in cardiac myocardium of SOD1 G93A and control mice. a , b Electron microscopy representative images of control SOD1 G93A myocardium. c , d ATP synthesis and oxygen consumption rate stimulated by pyruvate/malate in control (green column) and SOD1 G93A (red column) of isolated mitochondria. e Complex I activity in isolated mitochondria of control (green column) and of mutated muscles (red column) studied by the FeCN reduction in the presence of NADH. Data are expressed as mean ± SD, n = 3 for each group. Student t test for unpaired data was used for statistical evaluation
    Figure Legend Snippet: Mitochondrial ultrastructure and bioenergetics activity in cardiac myocardium of SOD1 G93A and control mice. a , b Electron microscopy representative images of control SOD1 G93A myocardium. c , d ATP synthesis and oxygen consumption rate stimulated by pyruvate/malate in control (green column) and SOD1 G93A (red column) of isolated mitochondria. e Complex I activity in isolated mitochondria of control (green column) and of mutated muscles (red column) studied by the FeCN reduction in the presence of NADH. Data are expressed as mean ± SD, n = 3 for each group. Student t test for unpaired data was used for statistical evaluation

    Techniques Used: Activity Assay, Mouse Assay, Electron Microscopy, Isolation

    8) Product Images from "Oligo-carrageenan kappa increases NADPH, ascorbate and glutathione syntheses and TRR/TRX activities enhancing photosynthesis, basal metabolism, and growth in Eucalyptus trees"

    Article Title: Oligo-carrageenan kappa increases NADPH, ascorbate and glutathione syntheses and TRR/TRX activities enhancing photosynthesis, basal metabolism, and growth in Eucalyptus trees

    Journal: Frontiers in Plant Science

    doi: 10.3389/fpls.2014.00512

    Effect of inhibitors on NADPH synthesis after treatment with OC kappa . Levels of NADPH (A) and NADH (C) in control Eucalyptus trees (empty circles), in trees treated with OC kappa (black circles) and cultivated for 0–4 months without additional treatment. Level of NADPH (B) and NADH (D) in control Eucalyptus trees (control), in trees treated with OC kappa (kappa) and in trees treated with CHS-828 and OC kappa (CHS+K), lycorine and OC kappa (Lyc+K), buthionine sulfoximine and OC kappa (BSO+K), and auranofin and OC kappa (Aur+K) and cultivated for 4 months without additional treatment. NAD(P)H levels are expressed in micrograms per gram of fresh tissue (FT). Symbols and bars represent mean values of three independent experiments and letters indicate significant differences ( p
    Figure Legend Snippet: Effect of inhibitors on NADPH synthesis after treatment with OC kappa . Levels of NADPH (A) and NADH (C) in control Eucalyptus trees (empty circles), in trees treated with OC kappa (black circles) and cultivated for 0–4 months without additional treatment. Level of NADPH (B) and NADH (D) in control Eucalyptus trees (control), in trees treated with OC kappa (kappa) and in trees treated with CHS-828 and OC kappa (CHS+K), lycorine and OC kappa (Lyc+K), buthionine sulfoximine and OC kappa (BSO+K), and auranofin and OC kappa (Aur+K) and cultivated for 4 months without additional treatment. NAD(P)H levels are expressed in micrograms per gram of fresh tissue (FT). Symbols and bars represent mean values of three independent experiments and letters indicate significant differences ( p

    Techniques Used:

    9) Product Images from "The PduQ Enzyme Is an Alcohol Dehydrogenase Used to Recycle NAD+ Internally within the Pdu Microcompartment of Salmonella enterica"

    Article Title: The PduQ Enzyme Is an Alcohol Dehydrogenase Used to Recycle NAD+ Internally within the Pdu Microcompartment of Salmonella enterica

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0047144

    Model for the Pdu microcompartment and the role of PduQ. The Pdu MCP consists of a protein shell that encapsulates enzymes and cofactors for metabolizing 1,2-propanediol. The shell is thought to be made from 9 proteins (PduABB'JKMNTU). Encapsulated within are enzymes for the activation of B 12 (III) to coenzyme B 12 (Ado-B 12 ) (PduS-O-GH) as well as three 1,2-PD degradative enzymes: Ado-B 12 -dependent diol dehydratase (PduCDE), propionaldehyde dehydrogenase (PduP) and 1-propanol dehydrogenase (PduQ). The proposed function of the Pdu MCP is to sequester propionaldehyde and channel it to downstream enzymes in order to prevent toxicity and DNA damage. The PduQ enzyme recycles NADH to NAD + internally within the MCP and this is needed for the activity of the PduP enzyme (this study). Alternatively, NADH can be recycled to NAD + by the electron transport chain although at a slower rate compared to internal recycling by PduQ. This latter process is thought to require movement of NAD + /NADH through specific pores that span the protein shell.
    Figure Legend Snippet: Model for the Pdu microcompartment and the role of PduQ. The Pdu MCP consists of a protein shell that encapsulates enzymes and cofactors for metabolizing 1,2-propanediol. The shell is thought to be made from 9 proteins (PduABB'JKMNTU). Encapsulated within are enzymes for the activation of B 12 (III) to coenzyme B 12 (Ado-B 12 ) (PduS-O-GH) as well as three 1,2-PD degradative enzymes: Ado-B 12 -dependent diol dehydratase (PduCDE), propionaldehyde dehydrogenase (PduP) and 1-propanol dehydrogenase (PduQ). The proposed function of the Pdu MCP is to sequester propionaldehyde and channel it to downstream enzymes in order to prevent toxicity and DNA damage. The PduQ enzyme recycles NADH to NAD + internally within the MCP and this is needed for the activity of the PduP enzyme (this study). Alternatively, NADH can be recycled to NAD + by the electron transport chain although at a slower rate compared to internal recycling by PduQ. This latter process is thought to require movement of NAD + /NADH through specific pores that span the protein shell.

    Techniques Used: Activation Assay, Activity Assay

    PduQ regenerates NAD + for the PduP reaction in purified Pdu MCPs. The PduP activity of purified MCPs was measured by monitoring the absorbance increase at 232 nm due to propionyl-CoA formation. Limiting NAD + (40 µM) was added in the assays. The propionyl-CoA produced by wild-type MCPs and ΔpduQ MCPs, respectively, reached 62.3 and 27.6 µM in 8 min. Because the concentration of propionyl-CoA produced by the wild-type exceeded the amount of NAD + added to the reaction (the PduP reaction produces 1 NADH per propionyl-CoA formed) recycling of NADH to NAD + is indicated.
    Figure Legend Snippet: PduQ regenerates NAD + for the PduP reaction in purified Pdu MCPs. The PduP activity of purified MCPs was measured by monitoring the absorbance increase at 232 nm due to propionyl-CoA formation. Limiting NAD + (40 µM) was added in the assays. The propionyl-CoA produced by wild-type MCPs and ΔpduQ MCPs, respectively, reached 62.3 and 27.6 µM in 8 min. Because the concentration of propionyl-CoA produced by the wild-type exceeded the amount of NAD + added to the reaction (the PduP reaction produces 1 NADH per propionyl-CoA formed) recycling of NADH to NAD + is indicated.

    Techniques Used: Purification, Activity Assay, Produced, Concentration Assay

    10) Product Images from "Beneficiary effect of Commiphora mukul ethanolic extract against high fructose diet induced abnormalities in carbohydrate and lipid metabolism in wistar rats"

    Article Title: Beneficiary effect of Commiphora mukul ethanolic extract against high fructose diet induced abnormalities in carbohydrate and lipid metabolism in wistar rats

    Journal: Journal of Traditional and Complementary Medicine

    doi: 10.1016/j.jtcme.2017.05.007

    Change in glucose-6, phosphate dehydrogenase activity of C, C + CM, F and F + CM groups during the 60 days experimental period. Values are ± S.E., (n = 8 animals).
    Figure Legend Snippet: Change in glucose-6, phosphate dehydrogenase activity of C, C + CM, F and F + CM groups during the 60 days experimental period. Values are ± S.E., (n = 8 animals).

    Techniques Used: Activity Assay

    11) Product Images from "YHR150w and YDR479c encode peroxisomal integral membrane proteins involved in the regulation of peroxisome number, size, and distribution in Saccharomyces cerevisiae"

    Article Title: YHR150w and YDR479c encode peroxisomal integral membrane proteins involved in the regulation of peroxisome number, size, and distribution in Saccharomyces cerevisiae

    Journal: The Journal of Cell Biology

    doi: 10.1083/jcb.200210130

    Yhr150p-prA and Ydr479p-prA remain at constant levels during incubation of S. cerevisiae in oleic acid–containing medium. Cells were grown for 16 h in glucose-containing YPD medium and then transferred to, and incubated in, oleic acid–containing YPBO medium. Aliquots of cells were removed from the YPBO medium at the times indicated, and total cell lysates were prepared. Equal amounts of protein from the total cell lysates were analyzed by SDS-PAGE and immunoblotting to visualize the protein A fusions. Antibodies directed against glucose-6-phosphatase (G6PDH) were used to confirm the loading of equal protein in each lane.
    Figure Legend Snippet: Yhr150p-prA and Ydr479p-prA remain at constant levels during incubation of S. cerevisiae in oleic acid–containing medium. Cells were grown for 16 h in glucose-containing YPD medium and then transferred to, and incubated in, oleic acid–containing YPBO medium. Aliquots of cells were removed from the YPBO medium at the times indicated, and total cell lysates were prepared. Equal amounts of protein from the total cell lysates were analyzed by SDS-PAGE and immunoblotting to visualize the protein A fusions. Antibodies directed against glucose-6-phosphatase (G6PDH) were used to confirm the loading of equal protein in each lane.

    Techniques Used: Incubation, SDS Page

    12) Product Images from "Switching of metabolic programs in response to light availability is an essential function of the cyanobacterial circadian output pathway"

    Article Title: Switching of metabolic programs in response to light availability is an essential function of the cyanobacterial circadian output pathway

    Journal: eLife

    doi: 10.7554/eLife.23210

    Characterization of strains used for the reconstitution experiments. Enzyme activities of glycogen phosphorylase, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in wild type, the rpaA - and the rpaA - +enzymes strains. Strains were incubated in BG-11 with 100 μM IPTG for 12 hr in light and harvested for the assay 3 hr after exposure to darkness. Error bars display the standard error of the mean of two experiments. DOI: http://dx.doi.org/10.7554/eLife.23210.015
    Figure Legend Snippet: Characterization of strains used for the reconstitution experiments. Enzyme activities of glycogen phosphorylase, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase in wild type, the rpaA - and the rpaA - +enzymes strains. Strains were incubated in BG-11 with 100 μM IPTG for 12 hr in light and harvested for the assay 3 hr after exposure to darkness. Error bars display the standard error of the mean of two experiments. DOI: http://dx.doi.org/10.7554/eLife.23210.015

    Techniques Used: Incubation

    13) Product Images from "A Differential Redox Regulation of the Pathways Metabolizing Glyceraldehyde-3-Phosphate Tunes the Production of Reducing Power in the Cytosol of Plant Cells"

    Article Title: A Differential Redox Regulation of the Pathways Metabolizing Glyceraldehyde-3-Phosphate Tunes the Production of Reducing Power in the Cytosol of Plant Cells

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms14048073

    Inactivation of each triose-P DHase by H 2 O 2 in the presence of substrates. (a) Incubation of np -Ga3PDHase (100% activity of 21 U/mg) with 1 mM H 2 O 2 in the presence of none (Δ), 1 mM Ga3P (□) or 100 μM NADP + (◇). Oxidation was followed in time during 20 min at 25 °C; ( b ) Incubation of Ga3PDHase (100% of activity of 38 U/mg) with 0.5 mM H 2 O 2 in the presence of none (▲), 1 mM NAD + (●), 1.2 mM Ga3P (▼) or 1 mM NAD + plus 1.2 mM Ga3P (■). Oxidation was followed in time during 40 s at 25 °C. Error bars represent the values deviation in three independent assays.
    Figure Legend Snippet: Inactivation of each triose-P DHase by H 2 O 2 in the presence of substrates. (a) Incubation of np -Ga3PDHase (100% activity of 21 U/mg) with 1 mM H 2 O 2 in the presence of none (Δ), 1 mM Ga3P (□) or 100 μM NADP + (◇). Oxidation was followed in time during 20 min at 25 °C; ( b ) Incubation of Ga3PDHase (100% of activity of 38 U/mg) with 0.5 mM H 2 O 2 in the presence of none (▲), 1 mM NAD + (●), 1.2 mM Ga3P (▼) or 1 mM NAD + plus 1.2 mM Ga3P (■). Oxidation was followed in time during 40 s at 25 °C. Error bars represent the values deviation in three independent assays.

    Techniques Used: Incubation, Activity Assay

    14) Product Images from "Histone H1.2 is a substrate for denitrase, an activity that reduces nitrotyrosine immunoreactivity in proteins"

    Article Title: Histone H1.2 is a substrate for denitrase, an activity that reduces nitrotyrosine immunoreactivity in proteins

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    doi: 10.1073/pnas.1131756100

    A 2D membrane assay for denitrase. The soluble fraction ( Upper ) and the CHAPS solubilized fraction ( Lower ) from peroxynitrite-treated RAW 264.7 cells were subjected to 2D membrane assay for denitrase. A spot corresponding pI = 8.5, M r of 45 kDa (arrow head) was found to disappear after incubation with cell lysate. This spot was excised and found out to be mouse Histone H1.2.
    Figure Legend Snippet: A 2D membrane assay for denitrase. The soluble fraction ( Upper ) and the CHAPS solubilized fraction ( Lower ) from peroxynitrite-treated RAW 264.7 cells were subjected to 2D membrane assay for denitrase. A spot corresponding pI = 8.5, M r of 45 kDa (arrow head) was found to disappear after incubation with cell lysate. This spot was excised and found out to be mouse Histone H1.2.

    Techniques Used: Incubation

    A 1D membrane assay for denitrase. Purified recombinant human Histone H1.2 was nitrated and subjected to 1D membrane assay for denitrase. ( A ) CTL, membrane treated with control solution; L/R, membrane treated with cell lysate from LPS-treated RAW 264.7 cells; (Z), membrane detected by anti-nitrotyrosine from Zymed; (U), membrane detected by anti-nitrotyrosine from Upstate Biotechnology. Nitrotyrosine immunoreactivity was detected with two different anti-nitrotyrosine mAbs on the control membrane at the M r of Histone H1.2. These signals of nitration were almost abolished after incubation of the membranes with cell lysate from LPS-treated RAW 264.7 cells. ( B ) The signals of 1D membrane assay for denitrase were quantified densitometrically. ( C ) Normalization of signals by Histone H1 Ab. The membrane incubated with LPS-treated RAW 267.4 cell lysate showed less intensity of nitrotyrosine immunoreactivity but similar intensity for anti-Histone H1 Ab.
    Figure Legend Snippet: A 1D membrane assay for denitrase. Purified recombinant human Histone H1.2 was nitrated and subjected to 1D membrane assay for denitrase. ( A ) CTL, membrane treated with control solution; L/R, membrane treated with cell lysate from LPS-treated RAW 264.7 cells; (Z), membrane detected by anti-nitrotyrosine from Zymed; (U), membrane detected by anti-nitrotyrosine from Upstate Biotechnology. Nitrotyrosine immunoreactivity was detected with two different anti-nitrotyrosine mAbs on the control membrane at the M r of Histone H1.2. These signals of nitration were almost abolished after incubation of the membranes with cell lysate from LPS-treated RAW 264.7 cells. ( B ) The signals of 1D membrane assay for denitrase were quantified densitometrically. ( C ) Normalization of signals by Histone H1 Ab. The membrane incubated with LPS-treated RAW 267.4 cell lysate showed less intensity of nitrotyrosine immunoreactivity but similar intensity for anti-Histone H1 Ab.

    Techniques Used: Purification, Recombinant, CTL Assay, Nitration, Incubation

    Time course analysis of denitrase activity. ( A ) Each piece of membrane containing nitrated histone H1.2 was incubated with cell lysate from LPS-treated RAW 264.7 cells for various lengths of time. The signals for the 1D membrane assay are shown. ( B ) The signals were quantified densitometrically and plotted according to the incubation time. Note that the y axis is logarithmic. A decrease of signal in accordance with the incubation time was seen.
    Figure Legend Snippet: Time course analysis of denitrase activity. ( A ) Each piece of membrane containing nitrated histone H1.2 was incubated with cell lysate from LPS-treated RAW 264.7 cells for various lengths of time. The signals for the 1D membrane assay are shown. ( B ) The signals were quantified densitometrically and plotted according to the incubation time. Note that the y axis is logarithmic. A decrease of signal in accordance with the incubation time was seen.

    Techniques Used: Activity Assay, Incubation

    Substrate specificity and the effect of proteasome inhibitor. ( A ) Nitrated Cu/Zn SOD was subjected to the 1D membrane assay and compared to Histone H1.2. The nitrotyrosine immunoreactivity of Histone H1.2 (upper arrow) was abolished by incubation with cell lysate from LPS-treated RAW 264.7 cells, whereas the immunoreactivity of nitrated Cu/Zn SOD (lower arrow) was not. In addition, a proteasome inhibitor, lactacystin (L) did not suppress this effect of cell lysate. CTL, membrane treated with control solution; L/R, membrane treated with cell lysate from LPS-treated RAW 264.7 cells. ( B ) The signals were quantified densitometrically.
    Figure Legend Snippet: Substrate specificity and the effect of proteasome inhibitor. ( A ) Nitrated Cu/Zn SOD was subjected to the 1D membrane assay and compared to Histone H1.2. The nitrotyrosine immunoreactivity of Histone H1.2 (upper arrow) was abolished by incubation with cell lysate from LPS-treated RAW 264.7 cells, whereas the immunoreactivity of nitrated Cu/Zn SOD (lower arrow) was not. In addition, a proteasome inhibitor, lactacystin (L) did not suppress this effect of cell lysate. CTL, membrane treated with control solution; L/R, membrane treated with cell lysate from LPS-treated RAW 264.7 cells. ( B ) The signals were quantified densitometrically.

    Techniques Used: Incubation, CTL Assay

    15) Product Images from "A Doped Polyaniline Modified Electrode Amperometric Biosensor for Gluconic Acid Determination in Grapes"

    Article Title: A Doped Polyaniline Modified Electrode Amperometric Biosensor for Gluconic Acid Determination in Grapes

    Journal: Sensors (Basel, Switzerland)

    doi: 10.3390/s140611097

    Calibration curve of GK-6PGDH/PEDGE/PANI-PAAMPSA/SPCE biosensor in optimized condition. Insert: amperometric biosensor response obtained injecting gluconic acid solutions into the FIA system.
    Figure Legend Snippet: Calibration curve of GK-6PGDH/PEDGE/PANI-PAAMPSA/SPCE biosensor in optimized condition. Insert: amperometric biosensor response obtained injecting gluconic acid solutions into the FIA system.

    Techniques Used:

    Effect of ATP/NADP + at different concentrations (mM), for their concentration ratio of 1.2 on the amperometric response obtained injecting 0.1 mM gluconic acid solutions into the FIA system.
    Figure Legend Snippet: Effect of ATP/NADP + at different concentrations (mM), for their concentration ratio of 1.2 on the amperometric response obtained injecting 0.1 mM gluconic acid solutions into the FIA system.

    Techniques Used: Concentration Assay

    Operational stability of gluconic acid biosensors during 6 h working. Amperometric response to gluconic acid 0.1 mM (70 injections) in PBS, applied potential of 0.1 V vs. Ag/AgCl.
    Figure Legend Snippet: Operational stability of gluconic acid biosensors during 6 h working. Amperometric response to gluconic acid 0.1 mM (70 injections) in PBS, applied potential of 0.1 V vs. Ag/AgCl.

    Techniques Used:

    Enzymatic reactions involved for gluconic acid chronoamperometric determination by GK and 6PGDH.
    Figure Legend Snippet: Enzymatic reactions involved for gluconic acid chronoamperometric determination by GK and 6PGDH.

    Techniques Used:

    Effect of ATP/NADP + ratio on the amperometric response obtained for 0.1 mM gluconic acid solution.
    Figure Legend Snippet: Effect of ATP/NADP + ratio on the amperometric response obtained for 0.1 mM gluconic acid solution.

    Techniques Used:

    16) Product Images from "Thioredoxin Is Involved in U(VI) and Cr(VI) Reduction in Desulfovibrio desulfuricans G20 ▿ G20 ▿ †"

    Article Title: Thioredoxin Is Involved in U(VI) and Cr(VI) Reduction in Desulfovibrio desulfuricans G20 ▿ G20 ▿ †

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.00197-09

    Effect of the recombinant thioredoxin operon on growth of E. coli in the presence of U(VI) (A) and U(VI) reduction by growing cultures sampled after 48 h (B). Results are shown for E. coli transformed with pXLthio1 (▪) or the untransformed strain
    Figure Legend Snippet: Effect of the recombinant thioredoxin operon on growth of E. coli in the presence of U(VI) (A) and U(VI) reduction by growing cultures sampled after 48 h (B). Results are shown for E. coli transformed with pXLthio1 (▪) or the untransformed strain

    Techniques Used: Recombinant, Transformation Assay

    (A) Relative expression of genes in the thioredoxin operon of G20 (▪) and in the Thio1 mutant (□). Expression of the CRP gene (not shown) was given a relative value of 1. (B) The influence U(VI) in the growth medium on expression of genes
    Figure Legend Snippet: (A) Relative expression of genes in the thioredoxin operon of G20 (▪) and in the Thio1 mutant (□). Expression of the CRP gene (not shown) was given a relative value of 1. (B) The influence U(VI) in the growth medium on expression of genes

    Techniques Used: Expressing, Mutagenesis

    Time courses of enzyme assays with the purified oxidoreductase. Assays were carried out independently to measure different components. (A) NADPH oxidation in an assay containing 1 mM U(VI), the purified oxidoreductase, thioredoxin reductase, thioredoxin,
    Figure Legend Snippet: Time courses of enzyme assays with the purified oxidoreductase. Assays were carried out independently to measure different components. (A) NADPH oxidation in an assay containing 1 mM U(VI), the purified oxidoreductase, thioredoxin reductase, thioredoxin,

    Techniques Used: Purification

    17) Product Images from "Methanol fermentation increases the production of NAD(P)H-dependent chemicals in synthetic methylotrophic Escherichia coli"

    Article Title: Methanol fermentation increases the production of NAD(P)H-dependent chemicals in synthetic methylotrophic Escherichia coli

    Journal: Biotechnology for Biofuels

    doi: 10.1186/s13068-019-1356-4

    Methanol bioconversion for improved lysine synthesis in synthetic methylotrophic E. coli . Enzymes required for the assimilation of methanol into central metabolism are shown in blue: MDH methanol dehydrogenase, HPS 3-hexulose-6-phosphate synthase, and PHI 6-phospho-3-hexuloisomerase. The genes overexpressed in the lysine biosynthetic pathway are shown in green. The cofactor generation pathway was reconstructed by expressing POS5 from S. cerevisiae to convert extra NADH and generate NADPH, which is shown in red
    Figure Legend Snippet: Methanol bioconversion for improved lysine synthesis in synthetic methylotrophic E. coli . Enzymes required for the assimilation of methanol into central metabolism are shown in blue: MDH methanol dehydrogenase, HPS 3-hexulose-6-phosphate synthase, and PHI 6-phospho-3-hexuloisomerase. The genes overexpressed in the lysine biosynthetic pathway are shown in green. The cofactor generation pathway was reconstructed by expressing POS5 from S. cerevisiae to convert extra NADH and generate NADPH, which is shown in red

    Techniques Used: Expressing

    Improved lysine production in synthetic methylotrophic E. coli . a Methylotrophic E. coli was engineered for the bioconversion of methanol to improve lysine production. The extra NADH from methanol consumption was engineered to generate NADPH for lysine production. b Lysine production in the strains BL21/ΔfrmA-ML and BL21/ΔfrmA-ML-POS5 cultivated with and without methanol. c Methanol consumption in the strains BL21/ΔfrmA-ML and BL21/ΔfrmA-ML-POS5. d The intracellular NADH pools in the strains BL21/ΔfrmA-ML and BL21/ΔfrmA-ML-POS5 cultivated with and without methanol. e The intracellular NADPH pools in the strains BL21/ΔfrmA-ML-POS5 cultivated with 55 mM glucose and 50 mM methanol. f Lysine production from 13 C-methanol in the strains BL21/ΔfrmA-Mdh2-Hps-Phi, BL21/ΔfrmA-ML, and BL21/ΔfrmA-ML-POS5. Error bars indicate standard error of the mean ( n = 3)
    Figure Legend Snippet: Improved lysine production in synthetic methylotrophic E. coli . a Methylotrophic E. coli was engineered for the bioconversion of methanol to improve lysine production. The extra NADH from methanol consumption was engineered to generate NADPH for lysine production. b Lysine production in the strains BL21/ΔfrmA-ML and BL21/ΔfrmA-ML-POS5 cultivated with and without methanol. c Methanol consumption in the strains BL21/ΔfrmA-ML and BL21/ΔfrmA-ML-POS5. d The intracellular NADH pools in the strains BL21/ΔfrmA-ML and BL21/ΔfrmA-ML-POS5 cultivated with and without methanol. e The intracellular NADPH pools in the strains BL21/ΔfrmA-ML-POS5 cultivated with 55 mM glucose and 50 mM methanol. f Lysine production from 13 C-methanol in the strains BL21/ΔfrmA-Mdh2-Hps-Phi, BL21/ΔfrmA-ML, and BL21/ΔfrmA-ML-POS5. Error bars indicate standard error of the mean ( n = 3)

    Techniques Used:

    18) Product Images from "p53 promotes AKT and SP1-dependent metabolism through the pentose phosphate pathway that inhibits apoptosis in response to Nutlin-3a"

    Article Title: p53 promotes AKT and SP1-dependent metabolism through the pentose phosphate pathway that inhibits apoptosis in response to Nutlin-3a

    Journal: Journal of Molecular Cell Biology

    doi: 10.1093/jmcb/mjx051

    Nutlin inhibits or promotes PPP. ( A ) Cells were treated with vehicle or Nutlin (10 μM) and/or MK2206 (10 μM) for 24 h. Lysates were analyzed for NADPH. Average NADPH level from triplicate was presented with SD indicated. ( B ) Cells were transfected with control siRNA, TIGAR siRNA, or G6PD siRNA and then treated with vehicle or Nutlin (10 μM) for 24 h. Lysates were analyzed for NADPH. Average NADPH level from triplicate was presented with SD indicated (left panel). Lysates were also immunoblotted for the indicated proteins (right panel). ( C ) cells were treated with Nutlin for a 12-h period and then fluxed with D-[1,2- 13 C] glucose for 15 min. Metabolites were analyzed by LC–MS. Ribose-5-P- 13 C1 levels in vehicle-treated and Nutlin-treated cells were presented with SD indicated. There is no significant difference between vehicle and Nutlin in U2OS cells ( P = 0.22). There is significant difference between vehicle and Nutlin in MHM cells ( P = 0.008). ( D ) Metabolism of 1,2- 13 C 2 -glucose through PPP to generate NADPH and into ribose-5-P- 13 C1 is schematically presented.
    Figure Legend Snippet: Nutlin inhibits or promotes PPP. ( A ) Cells were treated with vehicle or Nutlin (10 μM) and/or MK2206 (10 μM) for 24 h. Lysates were analyzed for NADPH. Average NADPH level from triplicate was presented with SD indicated. ( B ) Cells were transfected with control siRNA, TIGAR siRNA, or G6PD siRNA and then treated with vehicle or Nutlin (10 μM) for 24 h. Lysates were analyzed for NADPH. Average NADPH level from triplicate was presented with SD indicated (left panel). Lysates were also immunoblotted for the indicated proteins (right panel). ( C ) cells were treated with Nutlin for a 12-h period and then fluxed with D-[1,2- 13 C] glucose for 15 min. Metabolites were analyzed by LC–MS. Ribose-5-P- 13 C1 levels in vehicle-treated and Nutlin-treated cells were presented with SD indicated. There is no significant difference between vehicle and Nutlin in U2OS cells ( P = 0.22). There is significant difference between vehicle and Nutlin in MHM cells ( P = 0.008). ( D ) Metabolism of 1,2- 13 C 2 -glucose through PPP to generate NADPH and into ribose-5-P- 13 C1 is schematically presented.

    Techniques Used: Transfection, Liquid Chromatography with Mass Spectroscopy

    19) Product Images from "Control of Insulin Secretion by Production of Reactive Oxygen Species: Study Performed in Pancreatic Islets from Fed and 48-Hour Fasted Wistar Rats"

    Article Title: Control of Insulin Secretion by Production of Reactive Oxygen Species: Study Performed in Pancreatic Islets from Fed and 48-Hour Fasted Wistar Rats

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0158166

    Metabolic pathways associated to NADPH production from glucose and leucine oxidation in pancreatic islets. The sites indicated are: the pentose phosphate pathway and reactions involving intermediates of the Krebs cycle. ACO, aconitase; CYT, cytosol; DHEA, dehydroepiandrosterone; G3P, glyceraldehyde-3-phosphate dehydrogenase; GDH, glutamate dehydrogenase; GPx, glutathione peroxidase; GR, glutathione reductase; GSH, reduced glutathione; GSSG, oxidized glutathione; IDH, isocitrate dehydrogenase; ME, malic enzyme; MIT, mitochondria; PDH, pyruvate dehydrogenase; PPP, pentose phosphate pathway; SOD1, superoxide dismutase 1; 2OG, 2-oxoglutarate; α-KIC, α-ketoisocaproic acid.
    Figure Legend Snippet: Metabolic pathways associated to NADPH production from glucose and leucine oxidation in pancreatic islets. The sites indicated are: the pentose phosphate pathway and reactions involving intermediates of the Krebs cycle. ACO, aconitase; CYT, cytosol; DHEA, dehydroepiandrosterone; G3P, glyceraldehyde-3-phosphate dehydrogenase; GDH, glutamate dehydrogenase; GPx, glutathione peroxidase; GR, glutathione reductase; GSH, reduced glutathione; GSSG, oxidized glutathione; IDH, isocitrate dehydrogenase; ME, malic enzyme; MIT, mitochondria; PDH, pyruvate dehydrogenase; PPP, pentose phosphate pathway; SOD1, superoxide dismutase 1; 2OG, 2-oxoglutarate; α-KIC, α-ketoisocaproic acid.

    Techniques Used:

    Changes of NADPH/NADP+ ratio in INS-1E cells cultivated in the presence of 20 mM leucine and 2.8 or 16.7 mM glucose. NADPH/NADP + ratio in INS-1E cells incubated for 60 minutes in the presence of 2.8 or 16.7 mM glucose with or without addition of leucine (LEU—20 mM)–experimental protocol 6. The results are presented as mean ± SEM of three different cell preparations for each group. *p
    Figure Legend Snippet: Changes of NADPH/NADP+ ratio in INS-1E cells cultivated in the presence of 20 mM leucine and 2.8 or 16.7 mM glucose. NADPH/NADP + ratio in INS-1E cells incubated for 60 minutes in the presence of 2.8 or 16.7 mM glucose with or without addition of leucine (LEU—20 mM)–experimental protocol 6. The results are presented as mean ± SEM of three different cell preparations for each group. *p

    Techniques Used: Incubation

    20) Product Images from "The Impacts of Phosphorus Deficiency on the Photosynthetic Electron Transport Chain 1The Impacts of Phosphorus Deficiency on the Photosynthetic Electron Transport Chain 1 [OPEN]"

    Article Title: The Impacts of Phosphorus Deficiency on the Photosynthetic Electron Transport Chain 1The Impacts of Phosphorus Deficiency on the Photosynthetic Electron Transport Chain 1 [OPEN]

    Journal: Plant Physiology

    doi: 10.1104/pp.17.01624

    Concentrations of NADP + , NADPH, ATP, P i (PO 4 3− ), and starch in the youngest, fully expanded barley leaves. A, Concentrations of NADP + in leaf tissue. B, Concentrations of NADPH in leaf tissue. C, Concentrations of ATP in light-exposed thylakoids. D, Concentrations of PO 4 3− in leaf tissue. E, Concentrations of PO 4 3− in chloroplasts isolated after a 10-h dark period. F, Concentrations of PO 4 3− in chloroplasts isolated after a 4-h light period. G, Concentrations of starch in leaf tissue. The results are means ± SE ( n = 4, each with more than four technical replicates), and different letters represent statistically significant changes ( P
    Figure Legend Snippet: Concentrations of NADP + , NADPH, ATP, P i (PO 4 3− ), and starch in the youngest, fully expanded barley leaves. A, Concentrations of NADP + in leaf tissue. B, Concentrations of NADPH in leaf tissue. C, Concentrations of ATP in light-exposed thylakoids. D, Concentrations of PO 4 3− in leaf tissue. E, Concentrations of PO 4 3− in chloroplasts isolated after a 10-h dark period. F, Concentrations of PO 4 3− in chloroplasts isolated after a 4-h light period. G, Concentrations of starch in leaf tissue. The results are means ± SE ( n = 4, each with more than four technical replicates), and different letters represent statistically significant changes ( P

    Techniques Used: Isolation

    21) Product Images from "Crosstalk between cellular compartments protects against proteotoxicity and extends lifespan"

    Article Title: Crosstalk between cellular compartments protects against proteotoxicity and extends lifespan

    Journal: Scientific Reports

    doi: 10.1038/srep28751

    Localized chaperone deficiency does not activate any of the key cellular stress responses. Transcript levels of (a) cytosolic, (b) ER and (c) mitochondrial target genes in the chaperone deficient strains, compared to the WT. UBC6 was used as a control. The measurement was performed in biological and technical triplicate. (d) NADP/NADPH ratio is decreased in the chaperone deficient mutants. Data are represented as mean ± SD from 3 independent cultures, each measured in duplicate. ***p
    Figure Legend Snippet: Localized chaperone deficiency does not activate any of the key cellular stress responses. Transcript levels of (a) cytosolic, (b) ER and (c) mitochondrial target genes in the chaperone deficient strains, compared to the WT. UBC6 was used as a control. The measurement was performed in biological and technical triplicate. (d) NADP/NADPH ratio is decreased in the chaperone deficient mutants. Data are represented as mean ± SD from 3 independent cultures, each measured in duplicate. ***p

    Techniques Used:

    22) Product Images from "Heat-induced longevity in budding yeast requires respiratory metabolism and glutathione recycling"

    Article Title: Heat-induced longevity in budding yeast requires respiratory metabolism and glutathione recycling

    Journal: Aging (Albany NY)

    doi: 10.18632/aging.101560

    Glutathione recycling is essential for the heat shock induced replicative lifespan extension. ( A ) NADP + /NADPH ratio decreased in the WT during heat shock, suggesting increased NADPH levels. However, the ratio was unaffected by heat shock in the petite strain, the HAP4 Δ strain, in the presence of CCCP, in the strain deficient in G6PD ( ZWF1 Δ), or in the strain carrying overexpression of SOD2 . ( B ) GSH levels were increased in the WT during heat shock. Heat shock did not induce the GSH levels in the petite, HAP4 Δ strain, WT in the presence of CCCP, in the strain deficient in G6PD ( ZWF1 Δ), or in the strain carrying overexpression of SOD2 . ( C ) Oxygen consumption was increased under heat shock in the GLR1 Δ strain. Oxygen uptake was measured polarographically using a Clark-type electrode equipped oxygraph. ( D ) Mitochondrially produced superoxide (measured by FACS as MitoSOX fluorescence in 10000 cells) was increased in the GLR1 Δ strain during heat shock. The results are presented as the percentage of MitoSOX fluorescence detected in WT strain at optimal growth temperature. ( E ) qPCR measurement of gene expression levels showed that the absence of GLR1 did not affect the heat shock induced metabolic changes, also observed in the WT. Color of the squares on the heat map corresponds to the mean value of the log fold change from three biological and three technical replicates. UBC6 was used for normalization. ( F ) NADP + /NADPH ratio remained unchanged during heat shock in the GLR1 Δ strain. ( G ) GSH levels decreased during heat shock in the GLR1 Δ strain. (H) Mean and maximum replicative lifespan (RLS) were decreased in GLR1 Δ following heat shock. RLS of the double mutant HAP4 Δ GLR1 Δ revealed that GLR1 is epistatic to HAP4 under the conditions of mild heat shock. The number of curated cells if 87 for GLR1 Δ, 107 for GLR1 Δ HS, 96 for HAP4 ΔG LR1 Δ and 99 for HAP4 Δ GLR1 Δ HS. P value is
    Figure Legend Snippet: Glutathione recycling is essential for the heat shock induced replicative lifespan extension. ( A ) NADP + /NADPH ratio decreased in the WT during heat shock, suggesting increased NADPH levels. However, the ratio was unaffected by heat shock in the petite strain, the HAP4 Δ strain, in the presence of CCCP, in the strain deficient in G6PD ( ZWF1 Δ), or in the strain carrying overexpression of SOD2 . ( B ) GSH levels were increased in the WT during heat shock. Heat shock did not induce the GSH levels in the petite, HAP4 Δ strain, WT in the presence of CCCP, in the strain deficient in G6PD ( ZWF1 Δ), or in the strain carrying overexpression of SOD2 . ( C ) Oxygen consumption was increased under heat shock in the GLR1 Δ strain. Oxygen uptake was measured polarographically using a Clark-type electrode equipped oxygraph. ( D ) Mitochondrially produced superoxide (measured by FACS as MitoSOX fluorescence in 10000 cells) was increased in the GLR1 Δ strain during heat shock. The results are presented as the percentage of MitoSOX fluorescence detected in WT strain at optimal growth temperature. ( E ) qPCR measurement of gene expression levels showed that the absence of GLR1 did not affect the heat shock induced metabolic changes, also observed in the WT. Color of the squares on the heat map corresponds to the mean value of the log fold change from three biological and three technical replicates. UBC6 was used for normalization. ( F ) NADP + /NADPH ratio remained unchanged during heat shock in the GLR1 Δ strain. ( G ) GSH levels decreased during heat shock in the GLR1 Δ strain. (H) Mean and maximum replicative lifespan (RLS) were decreased in GLR1 Δ following heat shock. RLS of the double mutant HAP4 Δ GLR1 Δ revealed that GLR1 is epistatic to HAP4 under the conditions of mild heat shock. The number of curated cells if 87 for GLR1 Δ, 107 for GLR1 Δ HS, 96 for HAP4 ΔG LR1 Δ and 99 for HAP4 Δ GLR1 Δ HS. P value is

    Techniques Used: Over Expression, Produced, FACS, Fluorescence, Real-time Polymerase Chain Reaction, Expressing, Mutagenesis

    Yap1p is required for the mild heat shock induced RLS extension. ( A ) Oxygen consumption was increased under heat shock in the YAP1 Δ strain. Oxygen uptake was measured polarographically using a Clark-type electrode equipped oxygraph. ( B ) Mitochondrially produced superoxide (measured by FACS as MitoSOX fluorescence in 10000 cells) was increased in the YAP1 Δ strain during heat shock. The results are presented as the percentage of MitoSOX fluorescence detected in WT strain at optimal growth temperature. ( C ) qPCR measurement of gene expression levels showed that the absence of YAP1 did not affect the heat shock induced metabolic changes observed in WT. ( D ) NADP + /NADPH ratio was decreased during heat shock in the YAP1 Δ strain. ( E ) GSH levels were not affected by heat shock in the YAP1 Δ strain. ( F ) Mean and maximum replicative lifespan (RLS) were decreased in YAP1 Δ already at OGT, and further decreased following heat shock. The number of curated cells is 74 for YAP1 Δ and 84 for YAP1 Δ HS. Unless otherwise stated, data in graphs are mean ± SEM from three biological and three technical replicates. *** P
    Figure Legend Snippet: Yap1p is required for the mild heat shock induced RLS extension. ( A ) Oxygen consumption was increased under heat shock in the YAP1 Δ strain. Oxygen uptake was measured polarographically using a Clark-type electrode equipped oxygraph. ( B ) Mitochondrially produced superoxide (measured by FACS as MitoSOX fluorescence in 10000 cells) was increased in the YAP1 Δ strain during heat shock. The results are presented as the percentage of MitoSOX fluorescence detected in WT strain at optimal growth temperature. ( C ) qPCR measurement of gene expression levels showed that the absence of YAP1 did not affect the heat shock induced metabolic changes observed in WT. ( D ) NADP + /NADPH ratio was decreased during heat shock in the YAP1 Δ strain. ( E ) GSH levels were not affected by heat shock in the YAP1 Δ strain. ( F ) Mean and maximum replicative lifespan (RLS) were decreased in YAP1 Δ already at OGT, and further decreased following heat shock. The number of curated cells is 74 for YAP1 Δ and 84 for YAP1 Δ HS. Unless otherwise stated, data in graphs are mean ± SEM from three biological and three technical replicates. *** P

    Techniques Used: Produced, FACS, Fluorescence, Real-time Polymerase Chain Reaction, Expressing

    23) Product Images from "Physiological and Genomic Features of a Novel Sulfur-Oxidizing Gammaproteobacterium Belonging to a Previously Uncultivated Symbiotic Lineage Isolated from a Hydrothermal Vent"

    Article Title: Physiological and Genomic Features of a Novel Sulfur-Oxidizing Gammaproteobacterium Belonging to a Previously Uncultivated Symbiotic Lineage Isolated from a Hydrothermal Vent

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0104959

    Predicted central metabolism of strain Hiromi 1. Red lines indicate conserved pathways in most of the publically accessible genomes of the chemolithoautotrophic Gammaproteobacteria . EC numbers are given on each enzymatic reaction. Light blue font indicates concentrations (per pmol 10 10 cells) of metabolites. Metabolites that were not targets of the metabolomic analysis were given neither concentrations nor N.D. N.D.; not detected. Pi, phosphate; PPi, pyrophosphate; Glc, α-D-glucose; αG6P, α-D-glucose-6-phosphate; G1P, α-D-glucose-1-phosphate; βG6P, β-D-glucose-1-phosphate; F6P, β-D-fructose-6-phophate; FBP, β-D-fructose-1,6-bisphosphate; GAP, glyceraldehyde-3-phosphate; GBP, glycerate-1,3-bisphosphate; G3P, glycerate-3-phosphate; G2P, glycerate-2-phosphate; PEP, phosphoenolpyruvate; E4P, erythrose-4-phosphate; acetyl-P, acetyl phosphate; DHAP, dihydroxyacetone phosphate; X5P, D-xylulose-5-phosphate; SBP, D-sedoheptulose-1,7-bisphosphate; S7P, D-sedoheptulose-7-phosphate; RuBP, ribulose-1,5-bisphosphate; Ru5P, ribulose-5-phophate; R5P, D-ribose-5-phosphate.
    Figure Legend Snippet: Predicted central metabolism of strain Hiromi 1. Red lines indicate conserved pathways in most of the publically accessible genomes of the chemolithoautotrophic Gammaproteobacteria . EC numbers are given on each enzymatic reaction. Light blue font indicates concentrations (per pmol 10 10 cells) of metabolites. Metabolites that were not targets of the metabolomic analysis were given neither concentrations nor N.D. N.D.; not detected. Pi, phosphate; PPi, pyrophosphate; Glc, α-D-glucose; αG6P, α-D-glucose-6-phosphate; G1P, α-D-glucose-1-phosphate; βG6P, β-D-glucose-1-phosphate; F6P, β-D-fructose-6-phophate; FBP, β-D-fructose-1,6-bisphosphate; GAP, glyceraldehyde-3-phosphate; GBP, glycerate-1,3-bisphosphate; G3P, glycerate-3-phosphate; G2P, glycerate-2-phosphate; PEP, phosphoenolpyruvate; E4P, erythrose-4-phosphate; acetyl-P, acetyl phosphate; DHAP, dihydroxyacetone phosphate; X5P, D-xylulose-5-phosphate; SBP, D-sedoheptulose-1,7-bisphosphate; S7P, D-sedoheptulose-7-phosphate; RuBP, ribulose-1,5-bisphosphate; Ru5P, ribulose-5-phophate; R5P, D-ribose-5-phosphate.

    Techniques Used: Gas Chromatography

    24) Product Images from "Monoclonal antibody disulfide reduction during manufacturing"

    Article Title: Monoclonal antibody disulfide reduction during manufacturing

    Journal: mAbs

    doi: 10.4161/mabs.24725

    Figure 4. Influence of Product on Reduction using Thioredoxin System. Intact antibody, as measured by % Main peak in the NR CE-SDS analysis as a function of time.
    Figure Legend Snippet: Figure 4. Influence of Product on Reduction using Thioredoxin System. Intact antibody, as measured by % Main peak in the NR CE-SDS analysis as a function of time.

    Techniques Used:

    25) Product Images from "Aldehyde Reduction by Cytochrome P450"

    Article Title: Aldehyde Reduction by Cytochrome P450

    Journal: Current protocols in toxicology / editorial board, Mahin D. Maines (editor-in-chief) ... [et al.]

    doi: 10.1002/0471140856.tx0437s48

    Fluorescence excitation and emission spectra of 9-anthracene-carboxylic acid and 9-hydroxymethyl-anthracene
    Figure Legend Snippet: Fluorescence excitation and emission spectra of 9-anthracene-carboxylic acid and 9-hydroxymethyl-anthracene

    Techniques Used: Fluorescence

    26) Product Images from "Effect of Honokiol on Cytochrome P450 and UDP-Glucuronosyltransferase Enzyme Activities in Human Liver Microsomes"

    Article Title: Effect of Honokiol on Cytochrome P450 and UDP-Glucuronosyltransferase Enzyme Activities in Human Liver Microsomes

    Journal: Molecules

    doi: 10.3390/molecules180910681

    Representative Dixon plots for the inhibitory effects of honokiol on UGT1A9-catalyzed propofolglucuronidation in pooled human liver microsomes (H161). Each symbol represents the substrate concentration: propofol, 5 μM (∇), 10 μM (◯), 20 μM (∆), 40 μM (☐). Each data point represents the mean of triplicate experiments.
    Figure Legend Snippet: Representative Dixon plots for the inhibitory effects of honokiol on UGT1A9-catalyzed propofolglucuronidation in pooled human liver microsomes (H161). Each symbol represents the substrate concentration: propofol, 5 μM (∇), 10 μM (◯), 20 μM (∆), 40 μM (☐). Each data point represents the mean of triplicate experiments.

    Techniques Used: Concentration Assay

    27) Product Images from "Corydaline Inhibits Multiple Cytochrome P450 and UDP-Glucuronosyltransferase Enzyme Activities in Human Liver Microsomes"

    Article Title: Corydaline Inhibits Multiple Cytochrome P450 and UDP-Glucuronosyltransferase Enzyme Activities in Human Liver Microsomes

    Journal: Molecules

    doi: 10.3390/molecules16086591

    Effects of corydaline on rates of ( a ) UGT1A1-catalyzed 17β-estradiol 3-glucuronidation and ( b ) UGT1A9-catalyzed propofol glucuronidation in pooled human liver microsomes (H161). Lines represent the functions determined by nonlinear regression based on mixed competitive-noncompetitive equation. Each symbol represents the corydaline concentration: (a) 0 μM (⬤), 30 μM (◯), 60 μM (▼), 90 μM (▽), 120 μM (■); (b) 0 μM (⬤), 10 μM (◯), 20 μM (▼), 50 μM (▽). Each data point represents the mean of triplicate experiments.
    Figure Legend Snippet: Effects of corydaline on rates of ( a ) UGT1A1-catalyzed 17β-estradiol 3-glucuronidation and ( b ) UGT1A9-catalyzed propofol glucuronidation in pooled human liver microsomes (H161). Lines represent the functions determined by nonlinear regression based on mixed competitive-noncompetitive equation. Each symbol represents the corydaline concentration: (a) 0 μM (⬤), 30 μM (◯), 60 μM (▼), 90 μM (▽), 120 μM (■); (b) 0 μM (⬤), 10 μM (◯), 20 μM (▼), 50 μM (▽). Each data point represents the mean of triplicate experiments.

    Techniques Used: Concentration Assay

    28) Product Images from "Biochemical characterisation of the recombinant peroxiredoxin (FhePrx) of the liver fluke, Fasciola hepatica"

    Article Title: Biochemical characterisation of the recombinant peroxiredoxin (FhePrx) of the liver fluke, Fasciola hepatica

    Journal: Febs Letters

    doi: 10.1016/j.febslet.2006.08.019

    Steady-state kinetics for FhePrx at varying concentrations of thioredoxin (A) and susceptibility of FhePrx to inactivation by H 2 O 2 (B). (A) The enzyme-normalized reciprocal initial velocities are plotted against the reciprocal molarities of hydrogen peroxide. Thioredoxin concentrations are 5 μm (diamond), 7.5 μm (square), 10 μm (triangle) and 20 μm (X). (B) The time course of NADPH (0.25 mM) oxidation by recombinant FhePrx coupled with E. coli thioredoxin (10 μM) and thioredoxin reductase (2 U) is shown for H 2 O 2 concentrations of 0 mM (plus sign), 0.5 mM (diamond), 1.0 mM (square), 2.5 mM (triangle), 5.0 mM (×), 7.5 mM (× with bar), and 10 mM (closed circle). (C) FhePrx is sensitive to over-oxidation. The reaction rates from (B) were plotted over time at the varying hydrogen peroxide concentrations, then slopes of this data were plotted against reciprocal substrate (hydrogen peroxide) concentrations.
    Figure Legend Snippet: Steady-state kinetics for FhePrx at varying concentrations of thioredoxin (A) and susceptibility of FhePrx to inactivation by H 2 O 2 (B). (A) The enzyme-normalized reciprocal initial velocities are plotted against the reciprocal molarities of hydrogen peroxide. Thioredoxin concentrations are 5 μm (diamond), 7.5 μm (square), 10 μm (triangle) and 20 μm (X). (B) The time course of NADPH (0.25 mM) oxidation by recombinant FhePrx coupled with E. coli thioredoxin (10 μM) and thioredoxin reductase (2 U) is shown for H 2 O 2 concentrations of 0 mM (plus sign), 0.5 mM (diamond), 1.0 mM (square), 2.5 mM (triangle), 5.0 mM (×), 7.5 mM (× with bar), and 10 mM (closed circle). (C) FhePrx is sensitive to over-oxidation. The reaction rates from (B) were plotted over time at the varying hydrogen peroxide concentrations, then slopes of this data were plotted against reciprocal substrate (hydrogen peroxide) concentrations.

    Techniques Used: Recombinant

    Enzymatic activity of recombinant FhePrx, FhePrx protects plasmid DNA from oxidative nicking (A) and specific activity determined by NADPH consumption assay (B). (A) Molecular weight markers (1), plasmid pGEM 3zf only (2), pGEM 3zf with Fe 3+ and DTT (3), pGEM 3zf, Fe 3+ , DTT and recombinant FhePrx [880 ng] (4), pGEM 3zf, Fe 3+ , Ascorbic acid and recombinant FhePrx [880 ng] (5), pGEM 3zf, Fe 3+, DTT, and recombinant F. hepatica cathepsin L [1.4 μg] (6). The nicked (n) and supercoiled (s.c) form of plasmid are indicated by arrows. (B) Specific activity of FhePrx determined by NADPH (0.25 mM) consumption. Hydrogen peroxide at a concentration of 0.25 mM was provided as substrate. FhePrx, recombinant FhePrx, TR, E. coli thioredoxin and Trx, E. coli thioredoxin reductase. Values are the means of three independent experiments.
    Figure Legend Snippet: Enzymatic activity of recombinant FhePrx, FhePrx protects plasmid DNA from oxidative nicking (A) and specific activity determined by NADPH consumption assay (B). (A) Molecular weight markers (1), plasmid pGEM 3zf only (2), pGEM 3zf with Fe 3+ and DTT (3), pGEM 3zf, Fe 3+ , DTT and recombinant FhePrx [880 ng] (4), pGEM 3zf, Fe 3+ , Ascorbic acid and recombinant FhePrx [880 ng] (5), pGEM 3zf, Fe 3+, DTT, and recombinant F. hepatica cathepsin L [1.4 μg] (6). The nicked (n) and supercoiled (s.c) form of plasmid are indicated by arrows. (B) Specific activity of FhePrx determined by NADPH (0.25 mM) consumption. Hydrogen peroxide at a concentration of 0.25 mM was provided as substrate. FhePrx, recombinant FhePrx, TR, E. coli thioredoxin and Trx, E. coli thioredoxin reductase. Values are the means of three independent experiments.

    Techniques Used: Activity Assay, Recombinant, Plasmid Preparation, Molecular Weight, Concentration Assay

    29) Product Images from "Downregulation of thioredoxin reductase 1 expression in the substantia nigra pars compacta of Parkinson's disease mice"

    Article Title: Downregulation of thioredoxin reductase 1 expression in the substantia nigra pars compacta of Parkinson's disease mice

    Journal: Neural Regeneration Research

    doi: 10.3969/j.issn.1673-5374.2013.35.002

    Effect of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) intraperitoneal injection on thioredoxin reductase 1 (TR1) expression and thioredoxin reductase (TR) activity in the midbrain of mice. Seven days after MPTP treatment, the midbrain of mice was dissected out and TR1 expression levels and TR activity were evaluated. (A) Decreased level of TR1 protein was observed in MPTP-treated mice by western blot analysis. Data are expressed as the ratio of the absorbance of the target gene to that of the GAPDH control. (B) TR1 mRNA level in the midbrain of mice was measured using real-time reverse transcription-PCR. A significant reduction in TR1 mRNA level was found in the MPTP-treated mice. Results are expressed as the ratio of the absorbance of the target gene to that of the GAPDH control. (C) TR activity in the midbrain of the mice was evaluated using a thioredoxin reductase assay kit. MPTP decreased TR activity in the mouse midbrain. Data are expressed as mean ± SEM, and there were six mice in each group. a P
    Figure Legend Snippet: Effect of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) intraperitoneal injection on thioredoxin reductase 1 (TR1) expression and thioredoxin reductase (TR) activity in the midbrain of mice. Seven days after MPTP treatment, the midbrain of mice was dissected out and TR1 expression levels and TR activity were evaluated. (A) Decreased level of TR1 protein was observed in MPTP-treated mice by western blot analysis. Data are expressed as the ratio of the absorbance of the target gene to that of the GAPDH control. (B) TR1 mRNA level in the midbrain of mice was measured using real-time reverse transcription-PCR. A significant reduction in TR1 mRNA level was found in the MPTP-treated mice. Results are expressed as the ratio of the absorbance of the target gene to that of the GAPDH control. (C) TR activity in the midbrain of the mice was evaluated using a thioredoxin reductase assay kit. MPTP decreased TR activity in the mouse midbrain. Data are expressed as mean ± SEM, and there were six mice in each group. a P

    Techniques Used: Injection, Expressing, Activity Assay, Mouse Assay, Western Blot, Polymerase Chain Reaction, Reductase Assay

    Effect of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) intraperitoneal injection on the number of thioredoxin reductase 1 (TR1)-positive cells in the substantia nigra pars compacta (SNc) of mice. (A–D) Dopaminergic neurons stained for TR1 are shown in the representative images. Scale bars: Normal group: A, × 100; B, × 400. MPTP group: C, × 100; D, × 400. Positive TR1 expression is visible as brown yellow staining (arrows). (E) The number of TR1-positive cells was reduced in the MPTP group compared with the normal group. Data are expressed as mean ± SEM, and there were six mice in each group. a P
    Figure Legend Snippet: Effect of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) intraperitoneal injection on the number of thioredoxin reductase 1 (TR1)-positive cells in the substantia nigra pars compacta (SNc) of mice. (A–D) Dopaminergic neurons stained for TR1 are shown in the representative images. Scale bars: Normal group: A, × 100; B, × 400. MPTP group: C, × 100; D, × 400. Positive TR1 expression is visible as brown yellow staining (arrows). (E) The number of TR1-positive cells was reduced in the MPTP group compared with the normal group. Data are expressed as mean ± SEM, and there were six mice in each group. a P

    Techniques Used: Injection, Mouse Assay, Staining, Expressing

    30) Product Images from "Combined inhibition of glycolysis, the pentose cycle, and thioredoxin metabolism selectively increases cytotoxicity and oxidative stress in human breast and prostate cancer"

    Article Title: Combined inhibition of glycolysis, the pentose cycle, and thioredoxin metabolism selectively increases cytotoxicity and oxidative stress in human breast and prostate cancer

    Journal: Redox Biology

    doi: 10.1016/j.redox.2014.12.001

    The pathways involving glucose and hydroperoxide metabolism believed to be involved with protection of cancer cells from metabolic oxidative stress (inhibitors of Trx and GSH metabolism are shown in italics). 2DG competes with glucose for uptake into the cells competitively inhibiting pyruvate production and the pentose cycle after glucose-6-phosphate-dehydrogenase (G6PD). DHEA inhibits G6PD. The GSH and Trx dependent systems participate in the detoxification of H 2 O 2 and organic hydroperoxides. NADPH is a source of reducing equivalents for the Trx/GSH-dependent systems. BSO inhibits glutamate cysteine ligase (γ-GCL) preventing glutathione synthesis. Auranofin is the inhibitor of thioredoxin reductase (TrxR), which reduces the oxidized Trx to the reduced form. These inhibitors were used alone and in combination to increase the cancer cell oxidative stress, resulting in cancer cell cytotoxicity.
    Figure Legend Snippet: The pathways involving glucose and hydroperoxide metabolism believed to be involved with protection of cancer cells from metabolic oxidative stress (inhibitors of Trx and GSH metabolism are shown in italics). 2DG competes with glucose for uptake into the cells competitively inhibiting pyruvate production and the pentose cycle after glucose-6-phosphate-dehydrogenase (G6PD). DHEA inhibits G6PD. The GSH and Trx dependent systems participate in the detoxification of H 2 O 2 and organic hydroperoxides. NADPH is a source of reducing equivalents for the Trx/GSH-dependent systems. BSO inhibits glutamate cysteine ligase (γ-GCL) preventing glutathione synthesis. Auranofin is the inhibitor of thioredoxin reductase (TrxR), which reduces the oxidized Trx to the reduced form. These inhibitors were used alone and in combination to increase the cancer cell oxidative stress, resulting in cancer cell cytotoxicity.

    Techniques Used:

    31) Product Images from "Characterization of Key Residues and Membrane Association Domains in RDH10"

    Article Title: Characterization of Key Residues and Membrane Association Domains in RDH10

    Journal: The Biochemical journal

    doi: 10.1042/BJ20080812

    Reduction of atRAL to atROL by wtRDH10
    Figure Legend Snippet: Reduction of atRAL to atROL by wtRDH10

    Techniques Used:

    32) Product Images from "Development of a Fluorescence-based Trypanosoma cruzi CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease"

    Article Title: Development of a Fluorescence-based Trypanosoma cruzi CYP51 Inhibition Assay for Effective Compound Triaging in Drug Discovery Programmes for Chagas Disease

    Journal: PLoS Neglected Tropical Diseases

    doi: 10.1371/journal.pntd.0004014

    a: The effect of solvent on T . cruzi CYP51 activity; b: Cytochrome C reductase activity of recombinant preparations.
    Figure Legend Snippet: a: The effect of solvent on T . cruzi CYP51 activity; b: Cytochrome C reductase activity of recombinant preparations.

    Techniques Used: Activity Assay, Recombinant

    Trends of control 1 (closed circles, solvent only control, 0% inhibition), control 2 (open circles, reaction in absence of bactosomes, 100% inhibition) and Z’ (grey squares) of representative high-throughput screening runs of T . cruzi CYP51 FLINT (a) and cytochrome c reductase absorbance (b) assays in 384 well format. Error bars are the standard deviations of 16 replicates for each control and plate.
    Figure Legend Snippet: Trends of control 1 (closed circles, solvent only control, 0% inhibition), control 2 (open circles, reaction in absence of bactosomes, 100% inhibition) and Z’ (grey squares) of representative high-throughput screening runs of T . cruzi CYP51 FLINT (a) and cytochrome c reductase absorbance (b) assays in 384 well format. Error bars are the standard deviations of 16 replicates for each control and plate.

    Techniques Used: Inhibition, High Throughput Screening Assay

    33) Product Images from "NADPH accumulation is responsible for apoptosis in breast cancer cells induced by fatty acid synthase inhibition"

    Article Title: NADPH accumulation is responsible for apoptosis in breast cancer cells induced by fatty acid synthase inhibition

    Journal: Oncotarget

    doi: 10.18632/oncotarget.15936

    ROS is induced by FAS inhibition but not responsible for apoptosis ( A and B ) Cerulenin and orlistat induced ROS generation in MCF-7 and MDA-MB-231 cells. ROS levels were measured after cells were treated with FAS inhibitors as indicated for 2 h. ( C and D ) Effects of trolox, BHA and NAC on ROS generation in MCF7 and MDA-MB-231 cells induced by cerulenin and orlistat. ROS levels were measured after cells were treated with 10 μM of cerulenin or 100 μg/ml of orlistat in combination with 2 mM of trolox, 10 mM of NAC or 100 μM of BHA for 2 h. ( E and F ) Effects of trolox, BHA and NAC on apoptosis in MCF-7 and MDA-MB-231 cells induced by cerulenin and orlistat. Apoptosis was measured after cells were treated with 10 μM of cerulenin or 100 μg/ml of orlistat for 48 h. Error bar indicates ± SE ( n = 3).* p
    Figure Legend Snippet: ROS is induced by FAS inhibition but not responsible for apoptosis ( A and B ) Cerulenin and orlistat induced ROS generation in MCF-7 and MDA-MB-231 cells. ROS levels were measured after cells were treated with FAS inhibitors as indicated for 2 h. ( C and D ) Effects of trolox, BHA and NAC on ROS generation in MCF7 and MDA-MB-231 cells induced by cerulenin and orlistat. ROS levels were measured after cells were treated with 10 μM of cerulenin or 100 μg/ml of orlistat in combination with 2 mM of trolox, 10 mM of NAC or 100 μM of BHA for 2 h. ( E and F ) Effects of trolox, BHA and NAC on apoptosis in MCF-7 and MDA-MB-231 cells induced by cerulenin and orlistat. Apoptosis was measured after cells were treated with 10 μM of cerulenin or 100 μg/ml of orlistat for 48 h. Error bar indicates ± SE ( n = 3).* p

    Techniques Used: Inhibition, Multiple Displacement Amplification

    NADPH accumulation is responsible for apoptosis ( A and B ) Effects of trolox, BHA, NAC and DPI on NADPH/NADP + ratio in MCF-7 and MDA-MB-231 cells induced by cerulenin or orlistat. NADPH/NADP + was measured after cells were treated with 10 μM of cerulenin or 100 μg/ml of orlistat in combination with 2 mM of trolox, 10 mM of NAC, 100 μM of BHA or 5 μM of DPI for 2 h. ( C and D ) Effects of DPI on ROS generation in MCF-7 and MDA-MB-231 cells induced by cerulenin and orlistat. ROS levels were measured after cells were treated with 10 μM of cerulenin or 100 μg/ml of orlistat in combination with 5μM of DPI for 2 h. ( E and F ) Effects of DPI on apoptosis in MCF-7 and MDA-MB-231 cells induced by cerulenin and orlistat. Apoptosis was measured after cells were treated with 10 μM of cerulenin or 100 μg/ml of orlistat in combination with 5 μM of DPI for 48 h. Error bar indicates ± SE ( n = 3). * p
    Figure Legend Snippet: NADPH accumulation is responsible for apoptosis ( A and B ) Effects of trolox, BHA, NAC and DPI on NADPH/NADP + ratio in MCF-7 and MDA-MB-231 cells induced by cerulenin or orlistat. NADPH/NADP + was measured after cells were treated with 10 μM of cerulenin or 100 μg/ml of orlistat in combination with 2 mM of trolox, 10 mM of NAC, 100 μM of BHA or 5 μM of DPI for 2 h. ( C and D ) Effects of DPI on ROS generation in MCF-7 and MDA-MB-231 cells induced by cerulenin and orlistat. ROS levels were measured after cells were treated with 10 μM of cerulenin or 100 μg/ml of orlistat in combination with 5μM of DPI for 2 h. ( E and F ) Effects of DPI on apoptosis in MCF-7 and MDA-MB-231 cells induced by cerulenin and orlistat. Apoptosis was measured after cells were treated with 10 μM of cerulenin or 100 μg/ml of orlistat in combination with 5 μM of DPI for 48 h. Error bar indicates ± SE ( n = 3). * p

    Techniques Used: Multiple Displacement Amplification

    34) Product Images from "Mitochondrial NADP+-dependent isocitrate dehydrogenase deficiency increases cisplatin-induced oxidative damage in the kidney tubule cells"

    Article Title: Mitochondrial NADP+-dependent isocitrate dehydrogenase deficiency increases cisplatin-induced oxidative damage in the kidney tubule cells

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-018-0537-6

    Cisplatin administration decreases IDH2 expression and NADPH levels. Idh2 ‒/‒ mice ( h – j ) and wild-type ( Idh2 +/+ ) littermates ( a – j ) were intraperitoneally injected with either cisplatin (C, 20 mg/kg B.W.) or 0.9% saline (vehicle, V) once. Some mice were treated with Mito-T (M, 0.7 mg/kg B.W.) daily, beginning 7 days before cisplatin injection and continuing until experiments were completed. Kidneys were harvested at the indicated time after cisplatin injection. a BUN concentration was determined at the indicated time ( n = 4). b IDH2 and 4-HNE expression in mitochondrial fraction was determined by western blot analysis. c , d Band densities were measured by the ImageJ program. COX IV was used as loading control ( n = 4). e – g Activities of IDH2 ( e ), IDH1 ( f ), and IDH3 ( g ) were measured in mitochondria fraction ( e , g ) and cytosol fraction ( f ), respectively ( n = 4). h NADPH concentration in the kidney mitochondrial fraction was measured as described in the “Materials and methods” section. i Expression of IDH2 in mitochondrial fraction was detected by western blot analysis. j Band density was normalized to COX IV band using the ImageJ program. Results are expressed as means ± SE ( n = 3–4 per group). * p
    Figure Legend Snippet: Cisplatin administration decreases IDH2 expression and NADPH levels. Idh2 ‒/‒ mice ( h – j ) and wild-type ( Idh2 +/+ ) littermates ( a – j ) were intraperitoneally injected with either cisplatin (C, 20 mg/kg B.W.) or 0.9% saline (vehicle, V) once. Some mice were treated with Mito-T (M, 0.7 mg/kg B.W.) daily, beginning 7 days before cisplatin injection and continuing until experiments were completed. Kidneys were harvested at the indicated time after cisplatin injection. a BUN concentration was determined at the indicated time ( n = 4). b IDH2 and 4-HNE expression in mitochondrial fraction was determined by western blot analysis. c , d Band densities were measured by the ImageJ program. COX IV was used as loading control ( n = 4). e – g Activities of IDH2 ( e ), IDH1 ( f ), and IDH3 ( g ) were measured in mitochondria fraction ( e , g ) and cytosol fraction ( f ), respectively ( n = 4). h NADPH concentration in the kidney mitochondrial fraction was measured as described in the “Materials and methods” section. i Expression of IDH2 in mitochondrial fraction was detected by western blot analysis. j Band density was normalized to COX IV band using the ImageJ program. Results are expressed as means ± SE ( n = 3–4 per group). * p

    Techniques Used: Expressing, Mouse Assay, Injection, Concentration Assay, Western Blot

    35) Product Images from "Anoxic metabolism and biochemical production in Pseudomonas putida F1 driven by a bioelectrochemical system"

    Article Title: Anoxic metabolism and biochemical production in Pseudomonas putida F1 driven by a bioelectrochemical system

    Journal: Biotechnology for Biofuels

    doi: 10.1186/s13068-016-0452-y

    Estimated flux distribution in P. putida F1 during glucose oxidation in the anode compartment of the BES reactors with K 3 [Fe(CN) 6 ] (numbers on top ) and [Co(bpy) 3 ](ClO 4 ) 2 (numbers on bottom ) as mediators, respectively. Solid lines represent measured fluxes and fluxes derived from mass balancing. Dashed lines highlight assumed fluxes, not directly deducible from mass balancing. PQQ pyrroloquinolinequinone, FAD flavin adenine dinucleotide, UQH2 reduced ubiquinones, Cyt C cytochromes C, ADP adenosine diphosphate, ATP adenosine triphosphate, NADP + /NADPH nicotinamide adenine dinucleotide phosphate (oxidised / reduced), NAD + /NADH nicotinamide adenine dinucleotide (oxidised / reduced), 2KGA 2-keto-gluconic acid, 2K6PG 2-keto-gluconic acid-6-phosphate, 6PGNT 6-phosphogluconic acid, KDPG 2-keto-3-deoxy-phosphogluconic acid, GAP glyceraldehyde-3-phosphate, F16BP fructose-1,6-bisphosphate, F6P fructose-6-phosphate, G6P glucose-6-phosphate, 1,3BPG 1,3-biphosphoglyceric acid, 3PG 3-phosphoglyceric acid, 2PG 2-phosphoglyceric acid, PEP phosphoenolpyruvic acid, PYR pyruvic acid, Med ox oxidised mediator, Med red reduced mediator
    Figure Legend Snippet: Estimated flux distribution in P. putida F1 during glucose oxidation in the anode compartment of the BES reactors with K 3 [Fe(CN) 6 ] (numbers on top ) and [Co(bpy) 3 ](ClO 4 ) 2 (numbers on bottom ) as mediators, respectively. Solid lines represent measured fluxes and fluxes derived from mass balancing. Dashed lines highlight assumed fluxes, not directly deducible from mass balancing. PQQ pyrroloquinolinequinone, FAD flavin adenine dinucleotide, UQH2 reduced ubiquinones, Cyt C cytochromes C, ADP adenosine diphosphate, ATP adenosine triphosphate, NADP + /NADPH nicotinamide adenine dinucleotide phosphate (oxidised / reduced), NAD + /NADH nicotinamide adenine dinucleotide (oxidised / reduced), 2KGA 2-keto-gluconic acid, 2K6PG 2-keto-gluconic acid-6-phosphate, 6PGNT 6-phosphogluconic acid, KDPG 2-keto-3-deoxy-phosphogluconic acid, GAP glyceraldehyde-3-phosphate, F16BP fructose-1,6-bisphosphate, F6P fructose-6-phosphate, G6P glucose-6-phosphate, 1,3BPG 1,3-biphosphoglyceric acid, 3PG 3-phosphoglyceric acid, 2PG 2-phosphoglyceric acid, PEP phosphoenolpyruvic acid, PYR pyruvic acid, Med ox oxidised mediator, Med red reduced mediator

    Techniques Used: Derivative Assay

    a Specific intracellular ATP concentration in P. putida F1 under anaerobic conditions in the absence or presence of K 3 Fe(CN) 6 and electrodes. b Adenylate energy charge under the same conditions (AEC = (ATP + 0.5×ADP)/(ATP + ADP + AMP))
    Figure Legend Snippet: a Specific intracellular ATP concentration in P. putida F1 under anaerobic conditions in the absence or presence of K 3 Fe(CN) 6 and electrodes. b Adenylate energy charge under the same conditions (AEC = (ATP + 0.5×ADP)/(ATP + ADP + AMP))

    Techniques Used: Concentration Assay

    36) Product Images from "In vitro inhibition of human cytochrome P450 enzymes by the novel atypical antipsychotic drug asenapine: a prediction of possible drug–drug interactions"

    Article Title: In vitro inhibition of human cytochrome P450 enzymes by the novel atypical antipsychotic drug asenapine: a prediction of possible drug–drug interactions

    Journal: Pharmacological Reports

    doi: 10.1007/s43440-020-00089-z

    The influence of asenapine on the activity of CYP2D6 measured as a rate of bufuralol 1′-hydroxylation. a Human liver microsomes ( K m = 5.1 µM, V max = 5.59 pmol/mg protein/min). B Human cDNA-expressed CYP2D6 (Supersomes CYP2D6) ( K m = 6.6 µM, V max = 4.1 pmol/pmol CYP/min). Each point represents the mean value of two independent analyses. V velocity of the reaction, I the concentration of the inhibitor (asenapine), S the concentration of the substrate (bufuralol). The K i values and mechanisms of inhibition are shown in Table 1 . Dixon’s plots (the main plots): the bufuralol concentration of 10 µM (■), 25 µM (▲), and 50 µM (▼). Lineweaver–Burk’s plots (inserts): control—no asenapine (✱); the asenapine concentration of 0.5 µM ( x ), 1 µM ( ○ ), 5 µM ( □ ), and 10 µM ( △ )
    Figure Legend Snippet: The influence of asenapine on the activity of CYP2D6 measured as a rate of bufuralol 1′-hydroxylation. a Human liver microsomes ( K m = 5.1 µM, V max = 5.59 pmol/mg protein/min). B Human cDNA-expressed CYP2D6 (Supersomes CYP2D6) ( K m = 6.6 µM, V max = 4.1 pmol/pmol CYP/min). Each point represents the mean value of two independent analyses. V velocity of the reaction, I the concentration of the inhibitor (asenapine), S the concentration of the substrate (bufuralol). The K i values and mechanisms of inhibition are shown in Table 1 . Dixon’s plots (the main plots): the bufuralol concentration of 10 µM (■), 25 µM (▲), and 50 µM (▼). Lineweaver–Burk’s plots (inserts): control—no asenapine (✱); the asenapine concentration of 0.5 µM ( x ), 1 µM ( ○ ), 5 µM ( □ ), and 10 µM ( △ )

    Techniques Used: Activity Assay, Concentration Assay, Inhibition

    The influence of asenapine on the activity of CYP2C9 ( a ) measured as a rate of diclofenac 4′-hydroxylation and CYP2C19 ( b ) measured as a rate of perazine N -demethylation. The enzyme activity was estimated in human liver microsomes ( K m = 136.8 µM, V max = 21.02 nmol/mg protein/min for diclofenac 4′-hydroxylation; K m = 199.9 µM; V max = 1.58 nmol/mg protein/min for perazine N -demethylation) and Supersomes CYP2C9 and CYP2C19 ( K m = 131.9 µM, V max = 213.6 pmol/pmol CYP/min for diclofenac 4′-hydroxylation; K m = 162.3 µM, V max = 39.4 pmol/pmol CYP/min for perazine N -demethylation). Each point represents the mean value of two independent analyses. V velocity of the reaction, I the concentration of the inhibitor (asenapine), S the concentration of the substrate (diclofenac or perazine). Dixon’s plots: a the diclofenac concentration of 5 µM (■), 10 µM (▲), and 25 µM (▼); b the perazine concentration of 50 µM (■), 100 µM (▲), and 200 µM (▼)
    Figure Legend Snippet: The influence of asenapine on the activity of CYP2C9 ( a ) measured as a rate of diclofenac 4′-hydroxylation and CYP2C19 ( b ) measured as a rate of perazine N -demethylation. The enzyme activity was estimated in human liver microsomes ( K m = 136.8 µM, V max = 21.02 nmol/mg protein/min for diclofenac 4′-hydroxylation; K m = 199.9 µM; V max = 1.58 nmol/mg protein/min for perazine N -demethylation) and Supersomes CYP2C9 and CYP2C19 ( K m = 131.9 µM, V max = 213.6 pmol/pmol CYP/min for diclofenac 4′-hydroxylation; K m = 162.3 µM, V max = 39.4 pmol/pmol CYP/min for perazine N -demethylation). Each point represents the mean value of two independent analyses. V velocity of the reaction, I the concentration of the inhibitor (asenapine), S the concentration of the substrate (diclofenac or perazine). Dixon’s plots: a the diclofenac concentration of 5 µM (■), 10 µM (▲), and 25 µM (▼); b the perazine concentration of 50 µM (■), 100 µM (▲), and 200 µM (▼)

    Techniques Used: Activity Assay, Concentration Assay

    The influence of asenapine on the activity of CYP3A4 measured as a rate of testosterone 6β-hydroxylation. a Human liver microsomes ( K m = 283 µM, V max = 72.9 pmol/mg protein/min). b Human cDNA-expressed CYP3A4 (Supersomes CYP3A4) ( K m = 428 µM, V max = 42.9 pmol/pmol CYP/min). Each point represents the mean value of two independent analyses. V velocity of the reaction, I the concentration of the inhibitor (asenapine), S the concentration of the substrate (testosterone). The K i values and mechanisms of inhibition are shown in Table 1 . Dixon’s plots (the main plots): the testosterone concentration of 50 µM (■), 100 µM (▲), and 200 µM (▼). Lineweaver–Burk’s plots (inserts): control—no asenapine (✱); the asenapine concentration of 0.5 µM ( x ), 1 µM ( ○ ), 5 µM ( □ ), and 10 µM ( △ )
    Figure Legend Snippet: The influence of asenapine on the activity of CYP3A4 measured as a rate of testosterone 6β-hydroxylation. a Human liver microsomes ( K m = 283 µM, V max = 72.9 pmol/mg protein/min). b Human cDNA-expressed CYP3A4 (Supersomes CYP3A4) ( K m = 428 µM, V max = 42.9 pmol/pmol CYP/min). Each point represents the mean value of two independent analyses. V velocity of the reaction, I the concentration of the inhibitor (asenapine), S the concentration of the substrate (testosterone). The K i values and mechanisms of inhibition are shown in Table 1 . Dixon’s plots (the main plots): the testosterone concentration of 50 µM (■), 100 µM (▲), and 200 µM (▼). Lineweaver–Burk’s plots (inserts): control—no asenapine (✱); the asenapine concentration of 0.5 µM ( x ), 1 µM ( ○ ), 5 µM ( □ ), and 10 µM ( △ )

    Techniques Used: Activity Assay, Concentration Assay, Inhibition

    The influence of asenapine on the activity of CYP1A2 measured as a rate of caffeine 3- N -demethylation. a Human liver microsomes ( K m = 709 µM, V max = 9.67 pmol/mg protein/min). b Human cDNA-expressed CYP1A2 (Supersomes CYP1A2) ( K m = 705 µM, V max = 1.92 pmol/pmol CYP/min). Each point represents the mean value of two independent analyses. V velocity of the reaction, I the concentration of the inhibitor (asenapine), S the concentration of the substrate (caffeine). The K i values and mechanisms of inhibition are shown in Table 1 . Dixon’s plots (the main plots): the caffeine concentration of 200 µM (■), 400 µM (▲), and 800 µM (▼). Lineweaver–Burk’s plots (inserts): control—no asenapine (✱); the asenapine concentration of 0.5 µM ( x ), 1 µM ( ○ ), 5 µM ( □ ), and 10 µM ( △ )
    Figure Legend Snippet: The influence of asenapine on the activity of CYP1A2 measured as a rate of caffeine 3- N -demethylation. a Human liver microsomes ( K m = 709 µM, V max = 9.67 pmol/mg protein/min). b Human cDNA-expressed CYP1A2 (Supersomes CYP1A2) ( K m = 705 µM, V max = 1.92 pmol/pmol CYP/min). Each point represents the mean value of two independent analyses. V velocity of the reaction, I the concentration of the inhibitor (asenapine), S the concentration of the substrate (caffeine). The K i values and mechanisms of inhibition are shown in Table 1 . Dixon’s plots (the main plots): the caffeine concentration of 200 µM (■), 400 µM (▲), and 800 µM (▼). Lineweaver–Burk’s plots (inserts): control—no asenapine (✱); the asenapine concentration of 0.5 µM ( x ), 1 µM ( ○ ), 5 µM ( □ ), and 10 µM ( △ )

    Techniques Used: Activity Assay, Concentration Assay, Inhibition

    37) Product Images from "Transition to 37°C reveals importance of NADPH in mitigating oxidative stress in stored RBCs"

    Article Title: Transition to 37°C reveals importance of NADPH in mitigating oxidative stress in stored RBCs

    Journal: JCI Insight

    doi: 10.1172/jci.insight.126376

    Effects of PIPA on physical properties, Prx2 dimerization, and NADPH content of RBCs stored at 4°C prior to challenge with copper/ascorbate treatment or warming to 37°C. ( A ) Osmotic fragility based on solution osmolality leading to 50% hemolysis ( n = 3); and ( B ) percent lysed erythrocytes ( n = 3) in untreated RBC samples (circles) or samples treated (triangles) with 15% PIPA solution (sodium phosphate, inosine, sodium pyruvate, and adenine), followed by copper/ascorbate treatment for 0 (Control), 4, 24, or 48 hours at 4°C. ( C–E ) Osmotic fragility based on solution osmolality leading to 50% hemolysis in untreated RBC samples (white), samples treated with 15% PIPA solution on day 0 (light gray), ( C ) samples treated during 1 hour with 15% PIPA solution after storage (dark gray) ( n = 3), ( D ) samples treated with 30% PIPA solution on day 0 (dark gray) ( n = 6), or ( E ) samples treated with 15% PIPA solution at week 0 and week 3 (dark gray) ( n = 6), and stored at 4°C for 0, 2, 4, or 6 weeks. ( F ) Percent lysed erythrocytes ( n = 6) in untreated RBC samples (white) or samples treated with 15% PIPA solution on day 0 (gray) and stored at 4°C for 0, 2, 4, or 6 weeks. ( G ) Total NADP(H) and ( H ) NADPH determined by enzymatic cycling in untreated RBC samples (white) or samples treated with 15% (light gray) or 30% (dark gray) PIPA solution on day 0 and stored at 4°C for 0, 2, 4, or 6 weeks, followed by 20 hours at 37°C ( n = 6). Box plots show median, 25th and 75th percentiles (box), and minimum/maximum values (whiskers). Bars show mean values. * P ≤ 0.05, repeated-measures 2-way ANOVA; # P ≤ 0.05, Mann-Whitney nonparametric test.
    Figure Legend Snippet: Effects of PIPA on physical properties, Prx2 dimerization, and NADPH content of RBCs stored at 4°C prior to challenge with copper/ascorbate treatment or warming to 37°C. ( A ) Osmotic fragility based on solution osmolality leading to 50% hemolysis ( n = 3); and ( B ) percent lysed erythrocytes ( n = 3) in untreated RBC samples (circles) or samples treated (triangles) with 15% PIPA solution (sodium phosphate, inosine, sodium pyruvate, and adenine), followed by copper/ascorbate treatment for 0 (Control), 4, 24, or 48 hours at 4°C. ( C–E ) Osmotic fragility based on solution osmolality leading to 50% hemolysis in untreated RBC samples (white), samples treated with 15% PIPA solution on day 0 (light gray), ( C ) samples treated during 1 hour with 15% PIPA solution after storage (dark gray) ( n = 3), ( D ) samples treated with 30% PIPA solution on day 0 (dark gray) ( n = 6), or ( E ) samples treated with 15% PIPA solution at week 0 and week 3 (dark gray) ( n = 6), and stored at 4°C for 0, 2, 4, or 6 weeks. ( F ) Percent lysed erythrocytes ( n = 6) in untreated RBC samples (white) or samples treated with 15% PIPA solution on day 0 (gray) and stored at 4°C for 0, 2, 4, or 6 weeks. ( G ) Total NADP(H) and ( H ) NADPH determined by enzymatic cycling in untreated RBC samples (white) or samples treated with 15% (light gray) or 30% (dark gray) PIPA solution on day 0 and stored at 4°C for 0, 2, 4, or 6 weeks, followed by 20 hours at 37°C ( n = 6). Box plots show median, 25th and 75th percentiles (box), and minimum/maximum values (whiskers). Bars show mean values. * P ≤ 0.05, repeated-measures 2-way ANOVA; # P ≤ 0.05, Mann-Whitney nonparametric test.

    Techniques Used: MANN-WHITNEY

    Effects of copper/ascorbate-induced oxidative stress on physical properties, Prx2 dimerization, and NADP(H) content of freshly processed RBCs. ( A ) H 2 O 2 formation by RBCs (5% hematocrit) during 24 hours at 4°C in the absence (Control) or presence of copper/ascorbate (Asc) in fresh blood samples ( n = 6). ( B ) Percent Prx2 dimerization ( n = 6) in fresh blood samples exposed to copper/ascorbate for 0 (Control), 1, 4, 24, or 48 hours. ( C ) Total NADP(H) and ( D ) NADPH by enzymatic cycling in fresh blood samples exposed to copper/ascorbate for 0, 1, 4, 24, or 48 hours at 4°C ( n = 3). ( E ) Osmotic fragility based on solution osmolality leading to 50% hemolysis ( n = 3) and ( F ) percent lysed erythrocytes ( n = 3) in fresh blood samples exposed to copper/ascorbate for 0, 4, 24, or 48 hours at 4°C. Box plots show median, 25th and 75th percentiles (box), and minimum/maximum values (whiskers). Bars show mean values. * P ≤ 0.05, repeated-measures 1-way ANOVA in comparison to control; # P ≤ 0.05, Mann-Whitney nonparametric test.
    Figure Legend Snippet: Effects of copper/ascorbate-induced oxidative stress on physical properties, Prx2 dimerization, and NADP(H) content of freshly processed RBCs. ( A ) H 2 O 2 formation by RBCs (5% hematocrit) during 24 hours at 4°C in the absence (Control) or presence of copper/ascorbate (Asc) in fresh blood samples ( n = 6). ( B ) Percent Prx2 dimerization ( n = 6) in fresh blood samples exposed to copper/ascorbate for 0 (Control), 1, 4, 24, or 48 hours. ( C ) Total NADP(H) and ( D ) NADPH by enzymatic cycling in fresh blood samples exposed to copper/ascorbate for 0, 1, 4, 24, or 48 hours at 4°C ( n = 3). ( E ) Osmotic fragility based on solution osmolality leading to 50% hemolysis ( n = 3) and ( F ) percent lysed erythrocytes ( n = 3) in fresh blood samples exposed to copper/ascorbate for 0, 4, 24, or 48 hours at 4°C. Box plots show median, 25th and 75th percentiles (box), and minimum/maximum values (whiskers). Bars show mean values. * P ≤ 0.05, repeated-measures 1-way ANOVA in comparison to control; # P ≤ 0.05, Mann-Whitney nonparametric test.

    Techniques Used: MANN-WHITNEY

    Effects of incubation at 37°C on physical properties, Prx2 dimerization, and NADP(H) content of RBCs previously stored at 4°C. ( A ) Osmotic fragility based on solution osmolality leading to 50% hemolysis ( n = 6); and ( B ) elongation index (measure of RBC deformability) ( n = 6) at 0, 2, 4, and 6 weeks of storage at 4°C before (white) and after warming to 37°C for 4 hours (light gray) or 20 hours (dark gray). ( C ) Hydrogen peroxide formation as described in Methods during 20 hours at 4°C or 37°C (5% hematocrit) after storage for 0, 2, 4, or 6 weeks at 4°C ( n = 6). ( D ) Percentage Prx2 dimerization ( n = 6) at 0, 2, 4, and 6 weeks of storage at 4°C before (white) and after warming to 37°C for 4 hours or 20 hours. ( E ) Total NADP(H) and ( F ) NADPH determined by enzymatic cycling at 0, 2, 4, and 6 weeks of storage at 4°C before and after warming to 37°C for 24 hours ( n = 6). Box plots show median, 25th and 75th percentiles (box), and minimum/maximum values (whiskers). * P and full uncut gels.
    Figure Legend Snippet: Effects of incubation at 37°C on physical properties, Prx2 dimerization, and NADP(H) content of RBCs previously stored at 4°C. ( A ) Osmotic fragility based on solution osmolality leading to 50% hemolysis ( n = 6); and ( B ) elongation index (measure of RBC deformability) ( n = 6) at 0, 2, 4, and 6 weeks of storage at 4°C before (white) and after warming to 37°C for 4 hours (light gray) or 20 hours (dark gray). ( C ) Hydrogen peroxide formation as described in Methods during 20 hours at 4°C or 37°C (5% hematocrit) after storage for 0, 2, 4, or 6 weeks at 4°C ( n = 6). ( D ) Percentage Prx2 dimerization ( n = 6) at 0, 2, 4, and 6 weeks of storage at 4°C before (white) and after warming to 37°C for 4 hours or 20 hours. ( E ) Total NADP(H) and ( F ) NADPH determined by enzymatic cycling at 0, 2, 4, and 6 weeks of storage at 4°C before and after warming to 37°C for 24 hours ( n = 6). Box plots show median, 25th and 75th percentiles (box), and minimum/maximum values (whiskers). * P and full uncut gels.

    Techniques Used: Incubation

    38) Product Images from "Catalytic mechanism and substrate specificity of the ?-subunit of the voltage-gated potassium (Kv) channel"

    Article Title: Catalytic mechanism and substrate specificity of the ?-subunit of the voltage-gated potassium (Kv) channel

    Journal: Biochemistry

    doi: 10.1021/bi800301b

    Kinetic isotope effects on single turnover reactions of Kvβ2 (A) Replacement of intrinsically bound NADPH with NADPD. The NADPH bound to Kvβ2 (black) was oxidized by incubating the protein with 1 mM 4-nitrobenzaldehyde which led to a progressive decline in absorbance of the protein at 360 nm (inset; red line), indicating oxidation of NADPH to NADP + . The reaction mixture was then dialyzed for 16-20 h at 4°C against 0.2 M potassium phosphate, pH7.4. The protein was then incubated with synthesized NADPH or NADPD for 30 min at 4°C and dialyzed for 16-20 h. Reloading with NADPH (blue) or NADPD (green) led to an increase in A 360 , indicating re-formation of the protein-nucleotide binary complex; Time-dependent decrease in A 360 of NADPH (B) and NADPD (C) -bound Kvβ2 incubated with the indicated concentrations of 4-NB; and (D) The values of the pseudo-first order rate constant ( k trans to the data (R 2 =0.96); solid line, NADPH; dotted line, NADPD.
    Figure Legend Snippet: Kinetic isotope effects on single turnover reactions of Kvβ2 (A) Replacement of intrinsically bound NADPH with NADPD. The NADPH bound to Kvβ2 (black) was oxidized by incubating the protein with 1 mM 4-nitrobenzaldehyde which led to a progressive decline in absorbance of the protein at 360 nm (inset; red line), indicating oxidation of NADPH to NADP + . The reaction mixture was then dialyzed for 16-20 h at 4°C against 0.2 M potassium phosphate, pH7.4. The protein was then incubated with synthesized NADPH or NADPD for 30 min at 4°C and dialyzed for 16-20 h. Reloading with NADPH (blue) or NADPD (green) led to an increase in A 360 , indicating re-formation of the protein-nucleotide binary complex; Time-dependent decrease in A 360 of NADPH (B) and NADPD (C) -bound Kvβ2 incubated with the indicated concentrations of 4-NB; and (D) The values of the pseudo-first order rate constant ( k trans to the data (R 2 =0.96); solid line, NADPH; dotted line, NADPD.

    Techniques Used: Incubation, Synthesized

    39) Product Images from "THE ROLE OF THE HIGHLY CONSERVED THREONINE IN CYTOCHROME P450 2E1: PREVENTION OF H2O2-INDUCED INACTIVATION DURING ELECTRON TRANSFER"

    Article Title: THE ROLE OF THE HIGHLY CONSERVED THREONINE IN CYTOCHROME P450 2E1: PREVENTION OF H2O2-INDUCED INACTIVATION DURING ELECTRON TRANSFER

    Journal: Biochemistry

    doi: 10.1021/bi4004843

    HPLC profiles for P450 2E1 T303A incubated with or without NADPH. P450 2E1 T303A was incubated with reductase for 10 min in the absence (A) or presence (B) of 1 mM NADPH, and in the presence of NADPH together with 200 μM 4MP (C). After incubation,
    Figure Legend Snippet: HPLC profiles for P450 2E1 T303A incubated with or without NADPH. P450 2E1 T303A was incubated with reductase for 10 min in the absence (A) or presence (B) of 1 mM NADPH, and in the presence of NADPH together with 200 μM 4MP (C). After incubation,

    Techniques Used: High Performance Liquid Chromatography, Incubation

    Effects of Antioxidants on the NADPH-Supported Auto-Inactivation of P4350 2E1 T303A
    Figure Legend Snippet: Effects of Antioxidants on the NADPH-Supported Auto-Inactivation of P4350 2E1 T303A

    Techniques Used:

    Time-dependent loss in the activity of wild-type P450 2B4 and the T302A mutant. Wild-type P450 2B4 (A) or P450 2B4 T302A (B) were incubated with reductase in the absence of NADPH (●), in the presence of 1 mM NADPH (□), and in the presence
    Figure Legend Snippet: Time-dependent loss in the activity of wild-type P450 2B4 and the T302A mutant. Wild-type P450 2B4 (A) or P450 2B4 T302A (B) were incubated with reductase in the absence of NADPH (●), in the presence of 1 mM NADPH (□), and in the presence

    Techniques Used: Activity Assay, Mutagenesis, Incubation

    Hydrogen peroxide formation during the incubation of cytochrome P450s 2E1 or 2E1 T303A with reductase in the presence of NADPH. A, H 2 O 2 formation by wild-type 2E1 (○) and by the T303A mutant (●)in the presence of 1 mM NADPH. B, The effect
    Figure Legend Snippet: Hydrogen peroxide formation during the incubation of cytochrome P450s 2E1 or 2E1 T303A with reductase in the presence of NADPH. A, H 2 O 2 formation by wild-type 2E1 (○) and by the T303A mutant (●)in the presence of 1 mM NADPH. B, The effect

    Techniques Used: Incubation, Mutagenesis

    Possible pathway for the formation of the heme intermediates of P450 2E1 T303A when incubated with reductase and NADPH.
    Figure Legend Snippet: Possible pathway for the formation of the heme intermediates of P450 2E1 T303A when incubated with reductase and NADPH.

    Techniques Used: Incubation

    Effect of hydrogen peroxide on the NADPH-dependent inactivation of P450s 2E1 and 2E1 T303A. Wild-type P450 2E1 (A) and the P450 T303A mutant (B) were incubated with reductase in the absence (●) or presence (□) of 1 mM NADPH. Hydrogen peroxide
    Figure Legend Snippet: Effect of hydrogen peroxide on the NADPH-dependent inactivation of P450s 2E1 and 2E1 T303A. Wild-type P450 2E1 (A) and the P450 T303A mutant (B) were incubated with reductase in the absence (●) or presence (□) of 1 mM NADPH. Hydrogen peroxide

    Techniques Used: Mutagenesis, Incubation

    Time-dependent loss in activity for P450 2E1 wild-type and the T303A mutant. P450 2E1 wild-type (A) or P450 2E1 T303A (B) were incubated with reductase in the absence of NADPH (●), in the presence of 1 mM NADPH (□) or in the presence of
    Figure Legend Snippet: Time-dependent loss in activity for P450 2E1 wild-type and the T303A mutant. P450 2E1 wild-type (A) or P450 2E1 T303A (B) were incubated with reductase in the absence of NADPH (●), in the presence of 1 mM NADPH (□) or in the presence of

    Techniques Used: Activity Assay, Mutagenesis, Incubation

    40) Product Images from "Energy Conservation Associated with Ethanol Formation from H2 and CO2 in Clostridium autoethanogenum Involving Electron Bifurcation"

    Article Title: Energy Conservation Associated with Ethanol Formation from H2 and CO2 in Clostridium autoethanogenum Involving Electron Bifurcation

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.00399-15

    Scheme of the metabolism of Clostridium autoethanogenum growing on H 2 and CO 2 at pH 5.3, assuming that only acetic acid is formed and that the methylene-H 4 F reductase involved is electron bifurcating and NAD specific. For explanations, see the text. Hyt, electron-bifurcating and NADP-dependent [FeFe]-hydrogenase; Fdh, formate dehydrogenase; Nfn, electron-bifurcating and ferredoxin-dependent transhydrogenase; Rnf, membrane-associated and energy-conserving reduced ferredoxin:NAD + oxidoreductase.
    Figure Legend Snippet: Scheme of the metabolism of Clostridium autoethanogenum growing on H 2 and CO 2 at pH 5.3, assuming that only acetic acid is formed and that the methylene-H 4 F reductase involved is electron bifurcating and NAD specific. For explanations, see the text. Hyt, electron-bifurcating and NADP-dependent [FeFe]-hydrogenase; Fdh, formate dehydrogenase; Nfn, electron-bifurcating and ferredoxin-dependent transhydrogenase; Rnf, membrane-associated and energy-conserving reduced ferredoxin:NAD + oxidoreductase.

    Techniques Used:

    Related Articles

    Immunohistochemistry:

    Article Title: The timing and location of GDNF expression determines enteric nervous system structure and function
    Article Snippet: .. For ChAT/NADPH-d staining, 2 cm of proximal duodenum were first stained using the NADPH-d method, and then by ChAT immunohistochemistry (1:10, AB144P, Chemicon). .. For GDNF injected mice, 3 cm small bowel and 1.5 cm colon segments were analyzed.

    Fluorescence:

    Article Title: Transition to 37°C reveals importance of NADPH in mitigating oxidative stress in stored RBCs
    Article Snippet: .. NADPH and NADP+ were detected by fluorescence (Ex = 340 nm, Em = 445 nm) and UV254nm , respectively, and quantified using authentic standards (NADPH, Sigma-Aldrich; and NADP+ ) and ChemStation (Agilent Technologies). ..

    Purification:

    Article Title: GlnR Activation Induces Peroxide Resistance in Mycobacterial Biofilms
    Article Snippet: .. Purified NADP (Sigma # 10128031001) and NADPH (Sigma # 10107824001) were used for standard curves, which were used for determination of the nucleotides in each sample. ..

    Incubation:

    Article Title: Catalytic mechanism and substrate specificity of the ?-subunit of the voltage-gated potassium (Kv) channel
    Article Snippet: .. For synthesis, alcohol dehydrogenase 2U (1.21 mg) was incubated with 2 mM NADP+ in 50 mM Tris-HCl (pH7.8) and 300 mM 2-propanol or 2-propanol- d 8 (Sigma Chemical Co) for 1 h at 40°C. ..

    Activity Assay:

    Article Title: A regulatory role for Sec tRNA[Ser]Sec in selenoprotein synthesis
    Article Snippet: .. The activity of TR was measured optically through the reduction of sulfhydryl groups by a timed reaction with recombinant Escherichia coli thioredoxin (Calbiochem), insulin (Sigma), and NADPH (Sigma; ). .. Cell lysates were partially purified by affinity chromatography on a 2′5′-ADP-sepharose (Pharmacia) column.

    Staining:

    Article Title: The timing and location of GDNF expression determines enteric nervous system structure and function
    Article Snippet: .. For ChAT/NADPH-d staining, 2 cm of proximal duodenum were first stained using the NADPH-d method, and then by ChAT immunohistochemistry (1:10, AB144P, Chemicon). .. For GDNF injected mice, 3 cm small bowel and 1.5 cm colon segments were analyzed.

    Recombinant:

    Article Title: A regulatory role for Sec tRNA[Ser]Sec in selenoprotein synthesis
    Article Snippet: .. The activity of TR was measured optically through the reduction of sulfhydryl groups by a timed reaction with recombinant Escherichia coli thioredoxin (Calbiochem), insulin (Sigma), and NADPH (Sigma; ). .. Cell lysates were partially purified by affinity chromatography on a 2′5′-ADP-sepharose (Pharmacia) column.

    Similar Products

  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Millipore nadp
    ]). α-Helices 4, 8, 10, and 11 are labeled for reference and colored in yellow (monomer A, helices 4, 10, and 11) and orange (monomer B and helix 8). The <t>NADP</t> + - and Ca 2+ -binding sites are expanded in the labeled sub-panels. Calcium is shown as a pink sphere. NADP + ].
    Nadp, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 217 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nadp/product/Millipore
    Average 99 stars, based on 217 article reviews
    Price from $9.99 to $1999.99
    nadp - by Bioz Stars, 2020-09
    99/100 stars
      Buy from Supplier

    89
    Millipore nadph oxidase inhibitor
    ET formation depends on actin polymerization and partially on <t>NADPH</t> oxidase. Coelomocytes were isolated from E . andrei and in vitro stimulated for 24 hours with either PMA or bacteria X . bovienii (BACT). Prior to stimulation with PMA/BACT, some cells were pretreated with inhibitors as specifically indicated below. After addition of Sytox orange, the amount of extDNA was evaluated spectrofluorometerically. Effects of a) cytochalasin D inhibitor (CYTO-D), b) cytochalasin B inhibitor (CYTO-B), and c) peptidylarginine deiminase inhibitor (PAD4) on intensity of ETs formation measured by release of extDNA. d)Measurement of respiratory burst (NBT reduction) of coelomocytes isolated from E . andrei and in vitro stimulated for 24 hours with either PMA or BACT.Prior to stimulation with PMA/BACT, some cells were pretreated with <t>DPI,</t> an inhibitor of NADPH oxidase. e) Effects of DPIon intensity of ETs formation measured by release of extDNA. Each experiment was repeated at least 3 times and in each of them 3 to 4 individuals were used. Mean+SD, data for unstimulated cells are expressed as 100% and marked with a horizontal line, different letters (e.g. a vs. b or A vs. B) indicate statistically significant differences between the groups at p
    Nadph Oxidase Inhibitor, supplied by Millipore, used in various techniques. Bioz Stars score: 89/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/nadph oxidase inhibitor/product/Millipore
    Average 89 stars, based on 11 article reviews
    Price from $9.99 to $1999.99
    nadph oxidase inhibitor - by Bioz Stars, 2020-09
    89/100 stars
      Buy from Supplier

    90
    Millipore mitochondrial nadp
    Levels of <t>NADPH,</t> activity of GR and GPx, and GSSG/tGSH in the mitochondria from <t>Idh2</t> +/+ and Idh2 −/− mice after I/R. Idh2 +/+ ( Idh2 WT) and Idh2 −/− ( Idh2 KO) mice were subjected to either 25 minutes of bilateral renal ischemia or sham surgery. (A) Kidney sections were subjected to immunohistochemical staining using an anti-NAPDH antibody. Images were obtained from the outer medulla. Upper panels are at low magnification, lower panels are at high magnification of the dash-lined rectangle in upper panel. Brown indicates NADPH-positive signal. (B) Activity of GR, (C) GSSG/tGSH, and (D) the activity of GPx in the mitochondrial fractions from kidneys were determined as described in the Concise Methods section. Results are expressed as the mean±SEM ( n =6). * P
    Mitochondrial Nadp, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/mitochondrial nadp/product/Millipore
    Average 90 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    mitochondrial nadp - by Bioz Stars, 2020-09
    90/100 stars
      Buy from Supplier

    Image Search Results


    ]). α-Helices 4, 8, 10, and 11 are labeled for reference and colored in yellow (monomer A, helices 4, 10, and 11) and orange (monomer B and helix 8). The NADP + - and Ca 2+ -binding sites are expanded in the labeled sub-panels. Calcium is shown as a pink sphere. NADP + ].

    Journal: The Biochemical journal

    Article Title: Inhibitor potency varies widely among tumor-relevant human isocitrate dehydrogenase 1 mutants

    doi: 10.1042/BCJ20180424

    Figure Lengend Snippet: ]). α-Helices 4, 8, 10, and 11 are labeled for reference and colored in yellow (monomer A, helices 4, 10, and 11) and orange (monomer B and helix 8). The NADP + - and Ca 2+ -binding sites are expanded in the labeled sub-panels. Calcium is shown as a pink sphere. NADP + ].

    Article Snippet: NADPH (tetrasodium salt) and NADP+ (disodium salt) were purchased from Calbiochem (San Diego, CA).

    Techniques: Labeling, Binding Assay

    NADP + RMSF in WT, R132Q, R132H, and R132L IDH1 simulations with Ca 2+ bound. The RMSF per atom is colored onto each atom for ( A ) WT IDH1 (gray), ( B ) R132H IDH1 (green), ( C ) R132Q IDH1 (orange), and ( D ) R132L IDH1 (blue). The NADP + atoms that fluctuate more than 7 Å are in red, while the atoms which fluctuation less than 4 Å are in blue. Gradations between the two are modulated with white. The orientation of NADP + in the crystal structures is shown.

    Journal: The Biochemical journal

    Article Title: Inhibitor potency varies widely among tumor-relevant human isocitrate dehydrogenase 1 mutants

    doi: 10.1042/BCJ20180424

    Figure Lengend Snippet: NADP + RMSF in WT, R132Q, R132H, and R132L IDH1 simulations with Ca 2+ bound. The RMSF per atom is colored onto each atom for ( A ) WT IDH1 (gray), ( B ) R132H IDH1 (green), ( C ) R132Q IDH1 (orange), and ( D ) R132L IDH1 (blue). The NADP + atoms that fluctuate more than 7 Å are in red, while the atoms which fluctuation less than 4 Å are in blue. Gradations between the two are modulated with white. The orientation of NADP + in the crystal structures is shown.

    Article Snippet: NADPH (tetrasodium salt) and NADP+ (disodium salt) were purchased from Calbiochem (San Diego, CA).

    Techniques:

    ET formation depends on actin polymerization and partially on NADPH oxidase. Coelomocytes were isolated from E . andrei and in vitro stimulated for 24 hours with either PMA or bacteria X . bovienii (BACT). Prior to stimulation with PMA/BACT, some cells were pretreated with inhibitors as specifically indicated below. After addition of Sytox orange, the amount of extDNA was evaluated spectrofluorometerically. Effects of a) cytochalasin D inhibitor (CYTO-D), b) cytochalasin B inhibitor (CYTO-B), and c) peptidylarginine deiminase inhibitor (PAD4) on intensity of ETs formation measured by release of extDNA. d)Measurement of respiratory burst (NBT reduction) of coelomocytes isolated from E . andrei and in vitro stimulated for 24 hours with either PMA or BACT.Prior to stimulation with PMA/BACT, some cells were pretreated with DPI, an inhibitor of NADPH oxidase. e) Effects of DPIon intensity of ETs formation measured by release of extDNA. Each experiment was repeated at least 3 times and in each of them 3 to 4 individuals were used. Mean+SD, data for unstimulated cells are expressed as 100% and marked with a horizontal line, different letters (e.g. a vs. b or A vs. B) indicate statistically significant differences between the groups at p

    Journal: PLoS ONE

    Article Title: Conservative Mechanisms of Extracellular Trap Formation by Annelida Eisenia andrei: Serine Protease Activity Requirement

    doi: 10.1371/journal.pone.0159031

    Figure Lengend Snippet: ET formation depends on actin polymerization and partially on NADPH oxidase. Coelomocytes were isolated from E . andrei and in vitro stimulated for 24 hours with either PMA or bacteria X . bovienii (BACT). Prior to stimulation with PMA/BACT, some cells were pretreated with inhibitors as specifically indicated below. After addition of Sytox orange, the amount of extDNA was evaluated spectrofluorometerically. Effects of a) cytochalasin D inhibitor (CYTO-D), b) cytochalasin B inhibitor (CYTO-B), and c) peptidylarginine deiminase inhibitor (PAD4) on intensity of ETs formation measured by release of extDNA. d)Measurement of respiratory burst (NBT reduction) of coelomocytes isolated from E . andrei and in vitro stimulated for 24 hours with either PMA or BACT.Prior to stimulation with PMA/BACT, some cells were pretreated with DPI, an inhibitor of NADPH oxidase. e) Effects of DPIon intensity of ETs formation measured by release of extDNA. Each experiment was repeated at least 3 times and in each of them 3 to 4 individuals were used. Mean+SD, data for unstimulated cells are expressed as 100% and marked with a horizontal line, different letters (e.g. a vs. b or A vs. B) indicate statistically significant differences between the groups at p

    Article Snippet: Inhibitor of NADPH oxidase To determine involvement of ROS in ET production, cells were pre-treated with NADPH oxidase inhibitor, diphenyleneiodonium (DPI, 5 and 50 μM, Calbiochem, San Diego, California) in the in vitro conditions [ ].

    Techniques: Isolation, In Vitro

    Levels of NADPH, activity of GR and GPx, and GSSG/tGSH in the mitochondria from Idh2 +/+ and Idh2 −/− mice after I/R. Idh2 +/+ ( Idh2 WT) and Idh2 −/− ( Idh2 KO) mice were subjected to either 25 minutes of bilateral renal ischemia or sham surgery. (A) Kidney sections were subjected to immunohistochemical staining using an anti-NAPDH antibody. Images were obtained from the outer medulla. Upper panels are at low magnification, lower panels are at high magnification of the dash-lined rectangle in upper panel. Brown indicates NADPH-positive signal. (B) Activity of GR, (C) GSSG/tGSH, and (D) the activity of GPx in the mitochondrial fractions from kidneys were determined as described in the Concise Methods section. Results are expressed as the mean±SEM ( n =6). * P

    Journal: Journal of the American Society of Nephrology : JASN

    Article Title: Mitochondrial NADP+-Dependent Isocitrate Dehydrogenase Deficiency Exacerbates Mitochondrial and Cell Damage after Kidney Ischemia-Reperfusion Injury

    doi: 10.1681/ASN.2016030349

    Figure Lengend Snippet: Levels of NADPH, activity of GR and GPx, and GSSG/tGSH in the mitochondria from Idh2 +/+ and Idh2 −/− mice after I/R. Idh2 +/+ ( Idh2 WT) and Idh2 −/− ( Idh2 KO) mice were subjected to either 25 minutes of bilateral renal ischemia or sham surgery. (A) Kidney sections were subjected to immunohistochemical staining using an anti-NAPDH antibody. Images were obtained from the outer medulla. Upper panels are at low magnification, lower panels are at high magnification of the dash-lined rectangle in upper panel. Brown indicates NADPH-positive signal. (B) Activity of GR, (C) GSSG/tGSH, and (D) the activity of GPx in the mitochondrial fractions from kidneys were determined as described in the Concise Methods section. Results are expressed as the mean±SEM ( n =6). * P

    Article Snippet: Western blotting was performed using antibodies directed against mitochondrial NADP+ -dependent IDH2, cytosolic NADP+ -dependent IDH1, MnSOD (Calbiochem, San Diego, CA), copper-zinc superoxide dismutase (CuZnSOD; Chemicon, Temecula, CA), Opa1 (BD Biosciences, San Jose, CA), Drp1 (Cell Signaling Technology, Danvers, MA), Fis1 (Sigma-Aldrich), Bax (5B7) (EMD Millipore, Billerica, MA), Bax (6A7) (Santa Cruz Biotechnology, Santa Cruz, CA), Bcl-2 (EMD Millipore), XIAP (AnaSpec, San Jose, CA), Bcl-xL (BD Biosciences), cleaved caspase-3 (Cell Signaling Technology), 4-HNE (Abcam, Inc., Cambridge, MA), G6PD (Cell Signaling Technology), catalase (Fitzgerald, Concord, MA), Ly6G (eBioscience, San Diego, CA), cytochrome c (BD Biosciences), COX-IV (Abcam, Inc.), β -actin (Sigma-Aldrich), and GAPDH (Novus Biologicals, Littleton, CO).

    Techniques: Activity Assay, Mouse Assay, Immunohistochemistry, Staining