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pa d2hgdh pet20b(+) plasmid harboring pa0317 gene (GenScript corporation)

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GenScript corporation pa d2hgdh pet20b(+) plasmid harboring pa0317 gene
Pa D2hgdh Pet20b(+) Plasmid Harboring Pa0317 Gene, supplied by GenScript corporation, 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/product/plasmid+pa+d2hgdh+pet20b/pmc10034468-153-7-15?v=GenScript+corporation
Average 90 stars, based on 1 article reviews

pa d2hgdh pet20b(+) plasmid harboring pa0317 gene - by Bioz Stars, 2026-07

90/100 stars

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Flavin reduction of Pa D2HGDH. Left panel , a stopped-flow trace of the reductive-half reaction of Pa D2HGDH showing incomplete flavin reduction of Pa D2HGDH with 40 mM d -malate, fit to Equation . Note the log time scale. The instrument dead time following the mixing of the enzyme with the substrate was 2.2 ms. The residuals of the data fitting are shown in the bottom panel . Right panel , absorption spectra of Pa D2HGDH showing incomplete flavin reduction with saturating concentrations of d -malate at 100 mM or D2HG at 0.5 mM. The assay was carried out in 25 mM Tris–Cl, pH 7.4, at 25 °C. D2HG, d -2-hydroxyglutarate; Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa .

Journal: The Journal of Biological Chemistry

Article Title: Uncovering Zn 2+ as a cofactor of FAD-dependent Pseudomonas aeruginosa PAO1 d -2-hydroxyglutarate dehydrogenase

doi: 10.1016/j.jbc.2023.103007

Figure Lengend Snippet: Flavin reduction of Pa D2HGDH. Left panel , a stopped-flow trace of the reductive-half reaction of Pa D2HGDH showing incomplete flavin reduction of Pa D2HGDH with 40 mM d -malate, fit to Equation . Note the log time scale. The instrument dead time following the mixing of the enzyme with the substrate was 2.2 ms. The residuals of the data fitting are shown in the bottom panel . Right panel , absorption spectra of Pa D2HGDH showing incomplete flavin reduction with saturating concentrations of d -malate at 100 mM or D2HG at 0.5 mM. The assay was carried out in 25 mM Tris–Cl, pH 7.4, at 25 °C. D2HG, d -2-hydroxyglutarate; Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa .

Article Snippet: The Pa D2HGDH pET20b(+) plasmid harboring the PA0317 gene was designed in-lab and purchased from GenScript.

Techniques:

Activity assay of Pa D2HGDH after the exogenous addition of metals

Journal: The Journal of Biological Chemistry

Article Title: Uncovering Zn 2+ as a cofactor of FAD-dependent Pseudomonas aeruginosa PAO1 d -2-hydroxyglutarate dehydrogenase

doi: 10.1016/j.jbc.2023.103007

Figure Lengend Snippet: Activity assay of Pa D2HGDH after the exogenous addition of metals

Article Snippet: The Pa D2HGDH pET20b(+) plasmid harboring the PA0317 gene was designed in-lab and purchased from GenScript.

Techniques: Activity Assay

Kinetic properties of the different forms of Pa D2HGDH <xref ref-type=

a " width="100%" height="100%">

Journal: The Journal of Biological Chemistry

Article Title: Uncovering Zn 2+ as a cofactor of FAD-dependent Pseudomonas aeruginosa PAO1 d -2-hydroxyglutarate dehydrogenase

doi: 10.1016/j.jbc.2023.103007

Figure Lengend Snippet: Kinetic properties of the different forms of Pa D2HGDH a

Article Snippet: The Pa D2HGDH pET20b(+) plasmid harboring the PA0317 gene was designed in-lab and purchased from GenScript.

Techniques:

Substrate-induced reduction of Pa D2HGDH after purification with 1 mM Zn 2+ . Flavin reduction of Pa D2HGDH was recorded for ∼60 μM E-Zn 2+ -bound FAD ( blue ) with 5.4 mM d -malate as substrate in 25 mM NaPO 4 , 1 mM ZnCl 2 , pH 7.4, and 25 °C ( pink ). Sodium dithionite was used as a chemical reducing agent to mark the end point of flavin reduction ( black ). Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa .

Journal: The Journal of Biological Chemistry

Article Title: Uncovering Zn 2+ as a cofactor of FAD-dependent Pseudomonas aeruginosa PAO1 d -2-hydroxyglutarate dehydrogenase

doi: 10.1016/j.jbc.2023.103007

Figure Lengend Snippet: Substrate-induced reduction of Pa D2HGDH after purification with 1 mM Zn 2+ . Flavin reduction of Pa D2HGDH was recorded for ∼60 μM E-Zn 2+ -bound FAD ( blue ) with 5.4 mM d -malate as substrate in 25 mM NaPO 4 , 1 mM ZnCl 2 , pH 7.4, and 25 °C ( pink ). Sodium dithionite was used as a chemical reducing agent to mark the end point of flavin reduction ( black ). Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa .

Article Snippet: The Pa D2HGDH pET20b(+) plasmid harboring the PA0317 gene was designed in-lab and purchased from GenScript.

Techniques: Purification

UV–visible absorption spectrum of Pa D2HGDH with and without Zn 2+ . The UV−visible absorption spectrum of Pa D2HGDH was recorded in 25 mM NaPO 4 after purification with 1 mM ZnCl 2 ( red ), treatment with 1 mM EDTA ( blue ), and treatment with 100 mM EDTA ( black ), at pH 7.4 and 25 °C. Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa .

Journal: The Journal of Biological Chemistry

Article Title: Uncovering Zn 2+ as a cofactor of FAD-dependent Pseudomonas aeruginosa PAO1 d -2-hydroxyglutarate dehydrogenase

doi: 10.1016/j.jbc.2023.103007

Figure Lengend Snippet: UV–visible absorption spectrum of Pa D2HGDH with and without Zn 2+ . The UV−visible absorption spectrum of Pa D2HGDH was recorded in 25 mM NaPO 4 after purification with 1 mM ZnCl 2 ( red ), treatment with 1 mM EDTA ( blue ), and treatment with 100 mM EDTA ( black ), at pH 7.4 and 25 °C. Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa .

Article Snippet: The Pa D2HGDH pET20b(+) plasmid harboring the PA0317 gene was designed in-lab and purchased from GenScript.

Techniques: Purification

Effects of Zn 2+ on the properties of Pa D2HGDH-bound FAD

Journal: The Journal of Biological Chemistry

Article Title: Uncovering Zn 2+ as a cofactor of FAD-dependent Pseudomonas aeruginosa PAO1 d -2-hydroxyglutarate dehydrogenase

doi: 10.1016/j.jbc.2023.103007

Figure Lengend Snippet: Effects of Zn 2+ on the properties of Pa D2HGDH-bound FAD

Article Snippet: The Pa D2HGDH pET20b(+) plasmid harboring the PA0317 gene was designed in-lab and purchased from GenScript.

Techniques:

Effect of pH on the UV–visible absorption spectra of the different enzyme species of Pa D2HGDH. The UV−visible absorption spectra of ( A ) E-Zn 2+ , ( B ) E (E-Zn 2+ 100 mM EDTA ), and ( C ) free FAD were recorded at various pH values adjusted with NaOH in 25 mM NaPO 4 /NaPP i at 18 °C. A , inset , shows a charge–transfer band around the 530 nm to 930 nm region. Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa .

Journal: The Journal of Biological Chemistry

Article Title: Uncovering Zn 2+ as a cofactor of FAD-dependent Pseudomonas aeruginosa PAO1 d -2-hydroxyglutarate dehydrogenase

doi: 10.1016/j.jbc.2023.103007

Figure Lengend Snippet: Effect of pH on the UV–visible absorption spectra of the different enzyme species of Pa D2HGDH. The UV−visible absorption spectra of ( A ) E-Zn 2+ , ( B ) E (E-Zn 2+ 100 mM EDTA ), and ( C ) free FAD were recorded at various pH values adjusted with NaOH in 25 mM NaPO 4 /NaPP i at 18 °C. A , inset , shows a charge–transfer band around the 530 nm to 930 nm region. Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa .

Article Snippet: The Pa D2HGDH pET20b(+) plasmid harboring the PA0317 gene was designed in-lab and purchased from GenScript.

Techniques:

Difference spectra of the effect of pH on the UV–visible absorption spectra of the different enzyme species of Pa D2HGDH. The UV−visible absorption difference spectra were obtained by subtracting the flavin species at pH ∼9 from all subsequent species for ( A ) E-Zn 2+ ; ( B ) E (E-Zn 2+ 100 mM EDTA ), and ( C ) free FAD recorded in 25 mM NaPO 4 /NaPP i at 18 °C. Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa .

Journal: The Journal of Biological Chemistry

Article Title: Uncovering Zn 2+ as a cofactor of FAD-dependent Pseudomonas aeruginosa PAO1 d -2-hydroxyglutarate dehydrogenase

doi: 10.1016/j.jbc.2023.103007

Figure Lengend Snippet: Difference spectra of the effect of pH on the UV–visible absorption spectra of the different enzyme species of Pa D2HGDH. The UV−visible absorption difference spectra were obtained by subtracting the flavin species at pH ∼9 from all subsequent species for ( A ) E-Zn 2+ ; ( B ) E (E-Zn 2+ 100 mM EDTA ), and ( C ) free FAD recorded in 25 mM NaPO 4 /NaPP i at 18 °C. Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa .

Article Snippet: The Pa D2HGDH pET20b(+) plasmid harboring the PA0317 gene was designed in-lab and purchased from GenScript.

Techniques:

Metal activation of metallo-apo Pa D2HGDH. Apparent steady-state kinetics of inactivated Pa D2HGDH with varying concentrations of chloride salts of activating divalent metals and fixed saturating concentrations of 100 mM d -malate and 1 mM PMS. Enzyme inactivation was obtained after Pa D2HGDH treatment with 100 mM EDTA: ( A ) Zn 2+ activation, ( B ) Co 2+ activation, ( C ) Cd 2+ activation, ( D ) Ni 2+ activation, and ( E ) Mn 2+ activation. Enzyme activity was measured by monitoring the initial oxygen consumption rates. The data were fit to Equation . The reaction was carried out in 25 mM NaPO 4 , 1 mM ZnCl 2 , pH 7.4, and 25 °C. The resulting data extracted from the fit of the plots are summarized in <xref ref-type=

Table 5 . Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa ; PMS, phenazine methosulfate. " width="100%" height="100%">

Journal: The Journal of Biological Chemistry

Article Title: Uncovering Zn 2+ as a cofactor of FAD-dependent Pseudomonas aeruginosa PAO1 d -2-hydroxyglutarate dehydrogenase

doi: 10.1016/j.jbc.2023.103007

Figure Lengend Snippet: Metal activation of metallo-apo Pa D2HGDH. Apparent steady-state kinetics of inactivated Pa D2HGDH with varying concentrations of chloride salts of activating divalent metals and fixed saturating concentrations of 100 mM d -malate and 1 mM PMS. Enzyme inactivation was obtained after Pa D2HGDH treatment with 100 mM EDTA: ( A ) Zn 2+ activation, ( B ) Co 2+ activation, ( C ) Cd 2+ activation, ( D ) Ni 2+ activation, and ( E ) Mn 2+ activation. Enzyme activity was measured by monitoring the initial oxygen consumption rates. The data were fit to Equation . The reaction was carried out in 25 mM NaPO 4 , 1 mM ZnCl 2 , pH 7.4, and 25 °C. The resulting data extracted from the fit of the plots are summarized in Table 5 . Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa ; PMS, phenazine methosulfate.

Article Snippet: The Pa D2HGDH pET20b(+) plasmid harboring the PA0317 gene was designed in-lab and purchased from GenScript.

Techniques: Activation Assay, Activity Assay

Electrostatic potential map of Pa D2HGDH. A , general electrostatic potential map of the Pa D2HGDH homology model (built with SWISS-MODEL using a putative dehydrogenase from Rhodopseudomonas palustris (Protein Data Bank code: 3PM9 ). B , active site of Pa D2HGDH showing electronegative binding pocket of Zn 2+ ( blue sphere ) with FAD ( yellow sticks ) and D2HG ( green sticks ). C , highly electronegative protein surface pockets suitable for nonspecific Zn 2+ binding to Pa D2HGDH. Red, electronegative region; blue , electropositive region; white , neutral region. Images were generated after structural analyses of the Pa D2HGDH homology model using UCSF Chimera software . Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa .

Journal: The Journal of Biological Chemistry

Article Title: Uncovering Zn 2+ as a cofactor of FAD-dependent Pseudomonas aeruginosa PAO1 d -2-hydroxyglutarate dehydrogenase

doi: 10.1016/j.jbc.2023.103007

Figure Lengend Snippet: Electrostatic potential map of Pa D2HGDH. A , general electrostatic potential map of the Pa D2HGDH homology model (built with SWISS-MODEL using a putative dehydrogenase from Rhodopseudomonas palustris (Protein Data Bank code: 3PM9 ). B , active site of Pa D2HGDH showing electronegative binding pocket of Zn 2+ ( blue sphere ) with FAD ( yellow sticks ) and D2HG ( green sticks ). C , highly electronegative protein surface pockets suitable for nonspecific Zn 2+ binding to Pa D2HGDH. Red, electronegative region; blue , electropositive region; white , neutral region. Images were generated after structural analyses of the Pa D2HGDH homology model using UCSF Chimera software . Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa .

Article Snippet: The Pa D2HGDH pET20b(+) plasmid harboring the PA0317 gene was designed in-lab and purchased from GenScript.

Techniques: Binding Assay, Generated, Software

Proposed residues for Zn 2+ coordination in Pa D2HGDH using human D2HGDH as reference . A , Active-site topology of human D2HGDH (PDB code: 6LPP ) with bound D2HG and Zn 2+ showing residues for Zn 2+ coordination. B , active-site topology of the homology model of Pa D2HGDH (built with SWISS-MODEL using a putative dehydrogenase from Rhodopseudomonas palustris [PDB code: 3PM9 ]). Images were generated after structural analyses of the protein files using UCSF Chimera software . Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa .

Journal: The Journal of Biological Chemistry

Article Title: Uncovering Zn 2+ as a cofactor of FAD-dependent Pseudomonas aeruginosa PAO1 d -2-hydroxyglutarate dehydrogenase

doi: 10.1016/j.jbc.2023.103007

Figure Lengend Snippet: Proposed residues for Zn 2+ coordination in Pa D2HGDH using human D2HGDH as reference . A , Active-site topology of human D2HGDH (PDB code: 6LPP ) with bound D2HG and Zn 2+ showing residues for Zn 2+ coordination. B , active-site topology of the homology model of Pa D2HGDH (built with SWISS-MODEL using a putative dehydrogenase from Rhodopseudomonas palustris [PDB code: 3PM9 ]). Images were generated after structural analyses of the protein files using UCSF Chimera software . Pa D2HGDH, D2HGDH from Pseudomonas aeruginosa .

Article Snippet: The Pa D2HGDH pET20b(+) plasmid harboring the PA0317 gene was designed in-lab and purchased from GenScript.

Techniques: Generated, Software

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