Journal: Journal of natural products

Article Title: The Marine Cyanobacterial Metabolite Gallinamide A is a Potent and Selective Inhibitor of Human Cathepsin L

doi: 10.1021/np400727r

Figure Lengend Snippet: IC 50 Values and Selectivity Indices of Gallinamide A ( 1 ) for Cathepsins L, V, B, and H.

Article Snippet: Gallinamide was tested for inhibition of human cathepsin V (100 ng/mL; R&D Systems),human cathepsin B (100 ng/mL; R&D Systems), and human cathepsin H (50 ng/mL; Athens Research and Technology).

Techniques:

Journal: BMC Neuroscience

Article Title: Extracellular matrix-associated gene expression in adult sensory neuron populations cultured on a laminin substrate

doi: 10.1186/1471-2202-14-15

Figure Lengend Snippet: 36 genes were expressed by both DRG neuron populations on microarrays

Article Snippet: Pre-designed Assays-On-DemandTM Taqman ® rat gene expression assays (Applied Biosystems) were used to amplify the target genes ( AdamTs1 – Rn00577887; CTSH – Rn00564052 ; Fn1 – Rn00569575; Icam1 – Rn00564227; Igtb1 – Rn00566727; Lamb1 – Rn001473698; Spp1 – Rn00563571; Plaur – Rn00569290; Plat – Rn00565767).

Techniques:

Journal: BMC Neuroscience

Article Title: Extracellular matrix-associated gene expression in adult sensory neuron populations cultured on a laminin substrate

doi: 10.1186/1471-2202-14-15

Figure Lengend Snippet: Differences in expression for IB4 + compared to IB4 - neurons at t=0 and t=24LN as determined by each of the different assays employed

Article Snippet: Pre-designed Assays-On-DemandTM Taqman ® rat gene expression assays (Applied Biosystems) were used to amplify the target genes ( AdamTs1 – Rn00577887; CTSH – Rn00564052 ; Fn1 – Rn00569575; Icam1 – Rn00564227; Igtb1 – Rn00566727; Lamb1 – Rn001473698; Spp1 – Rn00563571; Plaur – Rn00569290; Plat – Rn00565767).

Techniques: Expressing

Journal: BMC Neuroscience

Article Title: Extracellular matrix-associated gene expression in adult sensory neuron populations cultured on a laminin substrate

doi: 10.1186/1471-2202-14-15

Figure Lengend Snippet: Differences in expression for IB4 + compared to IB4 - neurons at t=0 and t=24 as determined by each of the different assays employed

Article Snippet: Pre-designed Assays-On-DemandTM Taqman ® rat gene expression assays (Applied Biosystems) were used to amplify the target genes ( AdamTs1 – Rn00577887; CTSH – Rn00564052 ; Fn1 – Rn00569575; Icam1 – Rn00564227; Igtb1 – Rn00566727; Lamb1 – Rn001473698; Spp1 – Rn00563571; Plaur – Rn00569290; Plat – Rn00565767).

Techniques: Expressing

Journal: BMC Neuroscience

Article Title: Extracellular matrix-associated gene expression in adult sensory neuron populations cultured on a laminin substrate

doi: 10.1186/1471-2202-14-15

Figure Lengend Snippet: Expression of OPN, CTSH, AdamTs1 and Icam. ICC was used to detect the expression of osteopontin ( A , C , green), AdamTs1 ( E , G , green), CTSH ( I , L , green), Icam ( M , O , green); merged images show βIII tubulin (red, B , D , F , H , J , L , N , P ) and IB4 binding neurons (blue) at t=0 and t=24LN. Densitometric analyses are presented in Supp Fig . Scale bar-A-L, 30 μm; M-P, 20 μm.

Article Snippet: Pre-designed Assays-On-DemandTM Taqman ® rat gene expression assays (Applied Biosystems) were used to amplify the target genes ( AdamTs1 – Rn00577887; CTSH – Rn00564052 ; Fn1 – Rn00569575; Icam1 – Rn00564227; Igtb1 – Rn00566727; Lamb1 – Rn001473698; Spp1 – Rn00563571; Plaur – Rn00569290; Plat – Rn00565767).

Techniques: Expressing, Binding Assay

Journal: BMC Neuroscience

Article Title: Extracellular matrix-associated gene expression in adult sensory neuron populations cultured on a laminin substrate

doi: 10.1186/1471-2202-14-15

Figure Lengend Snippet: Immunohistochemisty of selected proteins in adult rat DRG cryosections . IHC and confocal microscopy was used to investigate expression of proteins encoded by a number of genes noted to be expressed at robust levels in the dissociated neurons, including those that have not been previously described in DRG neurons. Panels A – AdamTs1; B - Emmprin (Basigin); C – CTGF; D – CTSH; E - Fibronectin; F – Icam; G – Integrin β1; H – Laminin; I – MT-MMP1 (MMP19); J – Osteopontin (Spp1); K – RT-1 (RT-AWs); L – Plaur; M – SPARC; N – tPA (Plat); O – Peripherin; P – p75 neurotrophin receptor. Scale bar – 50 μm.

Article Snippet: Pre-designed Assays-On-DemandTM Taqman ® rat gene expression assays (Applied Biosystems) were used to amplify the target genes ( AdamTs1 – Rn00577887; CTSH – Rn00564052 ; Fn1 – Rn00569575; Icam1 – Rn00564227; Igtb1 – Rn00566727; Lamb1 – Rn001473698; Spp1 – Rn00563571; Plaur – Rn00569290; Plat – Rn00565767).

Techniques: Confocal Microscopy, Expressing

Journal: BMC Neuroscience

Article Title: Extracellular matrix-associated gene expression in adult sensory neuron populations cultured on a laminin substrate

doi: 10.1186/1471-2202-14-15

Figure Lengend Snippet: List of antibodies employed in this study

Article Snippet: Pre-designed Assays-On-DemandTM Taqman ® rat gene expression assays (Applied Biosystems) were used to amplify the target genes ( AdamTs1 – Rn00577887; CTSH – Rn00564052 ; Fn1 – Rn00569575; Icam1 – Rn00564227; Igtb1 – Rn00566727; Lamb1 – Rn001473698; Spp1 – Rn00563571; Plaur – Rn00569290; Plat – Rn00565767).

Techniques:

Journal: PLoS ONE

Article Title: Crystal structures of human procathepsin H

doi: 10.1371/journal.pone.0200374

Figure Lengend Snippet: (A) Overall structure of procathepsin H. Left, the prodomain and the mature domain are colored in violet and cyan respectively. The mini-chain is colored in blue. The prodomain contains two conserved sequence motifs: ERFNIN motif (orange) and GNFD motif (green). The four disulfide bonds in the structure are highlighted in yellow.Right, the 2 F o - F c electron density map (top) and F o - F c omit map (bottom) of the mini-chain are displayed as the grey mesh at a contour level of 1.2 σ and 3 σ respectively. (B) Superposition of procathepsin H (cyan) and procathepsin L (violet). (C) Superposition of human procathepsin H (cyan) and mature porcine cathepsin H (wheat). The mini-chain of human procathepsin H is shown in blue and that of mature porcine cathepsin H is shown in orange. (D) Hydrogen bonding interactions between the β strand from prodomain (violet) and β strand from right subdomain (cyan). Backbones of the residues involved are shown as sticks. The distances between atoms are indicated by dashes. (E) Interactions between hydrophobic residues from prodomain (violet) and the mature domain (cyan). Sidechains of the residues involved are shown as sticks. (F) Hydrogen bonding interactions between the C-terminal linker of prodomain (violet) and core enzyme (cyan). Backbones and sidechains of the residues involved are shown as sticks. The distances between atoms are indicated by dashes.

Article Snippet: Human cathepsin H (NM_004390) was cloned into PCDNA3.1 vector purchased from Origene Inc. Human cathepsin H point mutants were generated via site-directed mutagenesis using the QuickChange II Kit (Agilent).

Techniques: Sequencing

Journal: PLoS ONE

Article Title: Crystal structures of human procathepsin H

doi: 10.1371/journal.pone.0200374

Figure Lengend Snippet: (A) Water-mediated hydrogen bond network between mini-chain (blue) and the mature domain (cyan) in procathepsin H. Backbones and sidechains of the residues involved, as well as NAG moieties, are shown as sticks. Water molecules are shown as red spheres. The distances between atoms are indicated by dashes. (B) Hydrogen bonding interactions between mini-chain (orange) and the mature domain (sand) in mature porcine cathepsin H. Residues involved and the NAG moiety are shown as sticks. Water molecule is shown as red sphere. The distances between atoms are indicated by dashes. (C) Comparison of the mini-chain binding in the active site cleft in procathepsin H and mature porcine cathepsin H. The mature domain of procathepsin H is represented as surface in cyan and the mini-chain as green sticks. The mature domain of mature porcine cathepsin H is shown with surface representation in wheat and mini-chain as green sticks. (D) Superposition of the active site of procathepsin H (cyan) and mature porcine cathepsin H (wheat). (E) Superposition of the active site from procathepsin H (cyan) and procathepsin L (pink). In parentheses are the residue numbers of procathepsin L. (F) The active site in procathepsin H covered by a group of hydrophobic residues Tyr72P, Leu73P and Trp74P from the prodomain (violet). Active site residues are shown as green sticks.

Article Snippet: Human cathepsin H (NM_004390) was cloned into PCDNA3.1 vector purchased from Origene Inc. Human cathepsin H point mutants were generated via site-directed mutagenesis using the QuickChange II Kit (Agilent).

Techniques: Binding Assay

Journal: PLoS ONE

Article Title: Crystal structures of human procathepsin H

doi: 10.1371/journal.pone.0200374

Figure Lengend Snippet: (A) Measurement of cathepsin H aminopeptidase activity in HEK293T cell lines overexpressing the indicated forms of cathepsin H. Activity is expressed as RFUs (relative fluorescent units) detected after cleavage of cathepsin H substrate. (B) Western blot analysis of cathepsin H and actin in HEK293T cell lines expressing the indicated forms of cathepsin H. Arrow indicates pro form and mature form of cathepsin H.

Article Snippet: Human cathepsin H (NM_004390) was cloned into PCDNA3.1 vector purchased from Origene Inc. Human cathepsin H point mutants were generated via site-directed mutagenesis using the QuickChange II Kit (Agilent).

Techniques: Activity Assay, Western Blot, Expressing

Journal: PLoS ONE

Article Title: Crystal structures of human procathepsin H

doi: 10.1371/journal.pone.0200374

Figure Lengend Snippet: A . Coomassie gel of proCTSH incubated for 3 hours in sodium acetate(NaAC) buffer at various pHs (4.5, 5.0, 5.5), MES buffer at various pHs (5.5, 6.0, 6.5), thermolysin activated CTSH (TH), human liver purified CTSH (HS), and protein ladder (LD). Arrow indicates mature CTSH product. B. Assessment of CTSH enzymatic activity from samples in (A) and thermolysin alone (T). Data shown is relative fluorescence units (RFUs) from excitation emission readings at 360/480nm. C . Coomassie gel of procathepsin H (PH), proCTSH activated with CTSL for various minutes (5, 20, 60), thermolysin activated CTSH(TH) and protein ladder (LD). Arrow indicates mature CTSH product. D . Assessment of R-AMC (cathepsin H substrate) cleavage from proCathepsinH (proCTSH), CTSL, thermolysin (Therm), proCTSH activated with CTSL (CTSL CTSH), and thermolysin activated CTSH (Therm CTSH). Data shown is relative fluorescence units (RFUs) from excitation emission readings at 360/480nm.

Article Snippet: Human cathepsin H (NM_004390) was cloned into PCDNA3.1 vector purchased from Origene Inc. Human cathepsin H point mutants were generated via site-directed mutagenesis using the QuickChange II Kit (Agilent).

Techniques: Incubation, Purification, Activity Assay, Fluorescence

Journal: PLoS ONE

Article Title: Crystal structures of human procathepsin H

doi: 10.1371/journal.pone.0200374

Figure Lengend Snippet: The snapshots of the mini-chain and mature domain complexes before simulations ( left ) and the overlay of the mini-chain conformations during the simulations ( right ) in systems (A) 8pch_1glyc, (B) 6czk_1glyc, and (C) 6czk_2glyc. The cathepsin H (orange) and the procathepsin H (blue) systems are distinguished by their colors. The glycans present are shown in stick representation. (D) The per-residue RMSF of the mainchain atoms in the mini-chain calculated from the simulations and the B-factors from the crystal structures averaged by residue. The three systems 8pch_1glyc (filled orange circle), 6czk_1glyc (empty blue circle), and 6czk_2glyc (filled blue circle) are shown in different styles. The B-factors for 8PCH (orange) and 6CZK (blue) are shown as lines. (E) The distribution of native (left) and total (right) contact numbers computed from the simulations. Two residues are identified as contacting residues when any of the heavy atoms from the two are within 4.5 Å. The total contact number is evaluated for all residue pairs between the mini-chain and the mature domain. The native contacts are the ones from the total contacts that are also present in the native crystal structure. Results from the three systems 8pch_1glyc (solid orange line), 6czk_1glyc (dashed blue line), and 6czk_2glyc (solid blue line) are shown.

Article Snippet: Human cathepsin H (NM_004390) was cloned into PCDNA3.1 vector purchased from Origene Inc. Human cathepsin H point mutants were generated via site-directed mutagenesis using the QuickChange II Kit (Agilent).

Techniques:

Journal: PLoS ONE

Article Title: The Expression and Activity of Cathepsins D, H and K in Asthmatic Airways

doi: 10.1371/journal.pone.0057245

Figure Lengend Snippet: ASM (n = 3), lung fibroblast (n = 3) and airway epithelial (n = 3). Predicted band size indicated by (−). Abbreviations CTSB = cathepsin B, CTSD = cathepsin D, CTSF = cathepsin F, CTSH = cathepsin H, CTSK = cathepsin K, CTSL = cathepsin L, CTSS = cathepsin S and ASM = airway smooth muscle.

Article Snippet: Sections were then treated to minimize non-specific background staining and incubated with primary antibodies, goat anti-human CTSD (R&D Systems, Minneapolis, USA) [0.05 μg/mL], polyclonal mouse anti-human CTSH (Abnova, Taipei, Taiwan) [0.625 μg/mL] and CTSK (Abcam, Cambridge, UK) [1 μg/mL].

Techniques: