scholarly journals Mapping the Type I Collagen-binding Site on Pigment Epithelium-derived Factor

2002 ◽  
Vol 277 (47) ◽  
pp. 45400-45407 ◽  
Author(s):  
Christina Meyer ◽  
Luigi Notari ◽  
S. Patricia Becerra
Keyword(s):  
Binding Site ◽  
Type I Collagen ◽  
Pigment Epithelium ◽  
Type I ◽  
Collagen Binding ◽  
Pigment Epithelium Derived Factor

scholarly journals Inhibition of MMP-2 gelatinolysis by targeting exodomain–substrate interactions

2007 ◽  
Vol 406 (1) ◽  
pp. 147-155 ◽  
Author(s):  
Xiaoping Xu ◽  
Zhihua Chen ◽  
Yao Wang ◽  
Lynda Bonewald ◽  
Bjorn Steffensen
Keyword(s):  
Binding Site ◽  
Type I Collagen ◽  
Matrix Metalloproteinase 2 ◽  
Type I ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Collagen Binding ◽  
Collagen Peptides ◽  
Collagen Chain ◽  
Hydrolysis Of

MMP-2 (matrix metalloproteinase 2) contains a CBD (collagen-binding domain), which is essential for positioning gelatin substrate molecules relative to the catalytic site for cleavage. Deletion of the CBD or disruption of CBD-mediated gelatin binding inhibits gelatinolysis by MMP-2. To identify CBD-binding sites on type I collagen and collagen peptides with the capacity to compete CBD binding of gelatin and thereby inhibit gelatinolysis by MMP-2, we screened a one-bead one-peptide combinatorial peptide library with recombinant CBD as bait. Analyses of sequences from the CBD-binding peptides pointed to residues 715–721 in human α1(I) collagen chain as a binding site for CBD. A peptide (P713) including this collagen segment was synthesized for analyses. In SPR (surface plasmon resonance) assays, the CBD and MMP-2E404A, a catalytically inactive MMP-2 mutant, both bound immobilized P713 in a concentration-dependent manner, but not a scrambled control peptide. Furthermore, P713 competed gelatin binding by the CBD and MMP-2E404A. In control assays, neither of the non-collagen binding alkylated CBD or MMP-2 with deletion of CBD (MMP-2ΔCBD) bound P713. Consistent with the exodomain functions of the CBD, P713 inhibited ∼90% of the MMP-2 gelatin cleavage, but less than 20% of the MMP-2 activity on a peptide substrate (NFF-1) which does not require the CBD for cleavage. Confirming the specificity of the inhibition, P713 did not alter MMP-2ΔCBD or MMP-8 activities. These experiments identified a CBD-binding site on type I collagen and demonstrated that a corresponding synthetic peptide can inhibit hydrolysis of type I and IV collagens by competing CBD-mediated gelatin binding to MMP-2.

Microstructure dependent binding of pigment epithelium derived factor (PEDF) to type I collagen fibrils

2017 ◽  
Vol 199 (2) ◽  
pp. 132-139 ◽  
Author(s):  
Meagan Cauble ◽  
Phillip Yang ◽  
Ulrich Baumann ◽  
Jan M. Gebauer ◽  
Bradford G. Orr ◽  
...  
Keyword(s):  
Type I Collagen ◽  
Pigment Epithelium ◽  
Collagen Fibrils ◽  
Type I ◽  
Pigment Epithelium Derived Factor

scholarly journals Binding of soluble type I collagen to fibroblasts: effects of thermal activation of ligand, ligand concentration, pinocytosis, and cytoskeletal modifiers.

1982 ◽  
Vol 95 (3) ◽  
pp. 747-751 ◽  
Author(s):  
B D Goldberg
Keyword(s):  
Binding Site ◽  
Binding Sites ◽  
Thermal Activation ◽  
Type I Collagen ◽  
Type I ◽  
Collagen Binding ◽  
Bacterial Collagenase ◽  
Ligand Interactions ◽  
Organizing Center ◽  
High Ligand

Efficient binding of native, soluble 125I-labeled type I rat collagen to mouse 3T3 fibroblast monolayers requires prior warming of the ligand to 35-37 degrees C for 10-30 min. Decreased binding at high ligand concentrations is ascribed to ligand-ligand interactions rather than to negative cooperativity. Addition of bacterial collagenase to monolayers labeled with the 125I-ligand releases a constant fraction (80%) of the bound ligand over a 2-h interval at 37 degrees C, indicating that little of the ligand becomes inaccessible by pinocytosis. Colchicine (10(-7) M) and vinblastine (5 X 10(-8) M) do not inhibit binding by morphologically intact monolayers. Cytochalasins and concanavalin A show dose-related inhibition of binding by intact monolayers that is due to a reduction in the number of available binding sites rather than to a change in binding site affinity. The collagen binding site on the fibroblast surface is proposed as an organizing center for the assembly of periodic type I collagen fibrils.

Identification of Bilirubin Binding Site in Type I Collagen

2013 ◽  
Vol 19 (4) ◽  
pp. 357-364 ◽  
Author(s):  
Nagarajan Usharani ◽  
Gladstone Christopher Jayakumar ◽  
Swarna V. Kanth ◽  
Jonnalagadda Raghava Rao ◽  
Bangaru Chandrasekaran ◽  
...  
Keyword(s):  
Binding Site ◽  
Type I Collagen ◽  
Type I ◽  
Bilirubin Binding

The C-terminus type I collagen is a major binding site for heparin

1986 ◽  
Vol 882 (1) ◽  
pp. 1-5 ◽  
Author(s):  
Katharyn M. Keller ◽  
John M. Keller ◽  
Klaus Kühn
Keyword(s):  
Binding Site ◽  
Type I Collagen ◽  
Type I ◽  
C Terminus

A nuclear factor 1 binding site mediates the transcriptional activation of a type I collagen promoter by transforming growth factor-β

Cell ◽  
1988 ◽  
Vol 52 (3) ◽  
pp. 405-414 ◽  
Author(s):  
Pellegrino Rossi ◽  
Gerard Karsenty ◽  
Anita B. Roberts ◽  
Nanette S. Roche ◽  
Michael B. Sporn ◽  
...  
Keyword(s):  
Growth Factor ◽  
Binding Site ◽  
Transcriptional Activation ◽  
Type I Collagen ◽  
Transforming Growth Factor ◽  
Transforming Growth Factor Β ◽  
Type I ◽  
Nuclear Factor ◽  
Nuclear Factor 1 ◽  
Collagen Promoter

scholarly journals Characterization of the Collagen-Binding S-Layer Protein CbsA of Lactobacillus crispatus

2000 ◽  
Vol 182 (22) ◽  
pp. 6440-6450 ◽  
Author(s):  
Jouko Sillanpää ◽  
Beatriz Martínez ◽  
Jenni Antikainen ◽  
Takahiro Toba ◽  
Nisse Kalkkinen ◽  
...  
Keyword(s):  
Amino Acids ◽  
Type I Collagen ◽  
Signal Sequence ◽  
Type I ◽  
Binding Region ◽  
Sequence Comparisons ◽  
Collagen Binding ◽  
Reading Frame ◽  
Lactobacillus Crispatus ◽  
C Terminus

The cbsA gene of Lactobacillus crispatusstrain JCM 5810, encoding a protein that mediates adhesiveness to collagens, was characterized and expressed in Escherichia coli. The cbsA open reading frame encoded a signal sequence of 30 amino acids and a mature polypeptide of 410 amino acids with typical features of a bacterial S-layer protein. ThecbsA gene product was expressed as a His tag fusion protein, purified by affinity chromatography, and shown to bind solubilized as well as immobilized type I and IV collagens. Three otherLactobacillus S-layer proteins, SlpA, CbsB, and SlpnB, bound collagens only weakly, and sequence comparisons of CbsA with these S-layer proteins were used to select sites in cbsAwhere deletions and mutations were introduced. In addition, hybrid S-layer proteins that contained the N or the C terminus from CbsA, SlpA, or SlpnB as well as N- and C-terminally truncated peptides from CbsA were constructed by gene fusion. Analysis of these molecules revealed the major collagen-binding region within the N-terminal 287 residues and a weaker type I collagen-binding region in the C terminus of the CbsA molecule. The mutated or hybrid CbsA molecules and peptides that failed to polymerize into a periodic S-layer did not bind collagens, suggesting that the crystal structure with a regular array is optimal for expression of collagen binding by CbsA. Strain JCM 5810 was found to contain another S-layer gene termed cbsB that was 44% identical in sequence to cbsA. RNA analysis showed that cbsA, but not cbsB, was transcribed under laboratory conditions. S-layer-protein-expressing cells of strain JCM 5810 adhered to collagen-containing regions in the chicken colon, suggesting that CbsA-mediated collagen binding represents a true tissue adherence property of L. crispatus.

Integrin alpha 2 I-domain is a binding site for collagens

1995 ◽  
Vol 108 (4) ◽  
pp. 1629-1637 ◽  
Author(s):  
D. Tuckwell ◽  
D.A. Calderwood ◽  
L.J. Green ◽  
M.J. Humphries
Keyword(s):  
Type I Collagen ◽  
Type I ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
Collagen Binding ◽  
Functional Antibodies ◽  
A Domains ◽  
Domain I ◽  
Von Willebrand ◽  
Willebrand Factor

Integrins alpha 1 beta 1 and alpha 2 beta 1 are major cellular receptors for collagens. The alpha 1 and alpha 2 subunits contain a approximately 200 amino acid inserted domain (I-domain) in their N-terminal region and, because of the homology between the I-domains and the collagen-binding A-domains of von Willebrand factor, it has been suggested that the I-domains might mediate the collagen-binding functions of alpha 1 beta 1 and alpha 2 beta 1. In order to fully investigate this hypothesis, we have generated recombinant human alpha 2 I-domain (r alpha 2I) by reverse transcriptase-polymerase chain reaction/bacterial expression and tested its ability to mediate the collagen-binding functions of alpha 2 beta 1. R alpha 2 I binds specifically to type I collagen in a concentration-dependent manner: binding is cation dependent and, like the complete receptor, is supported by magnesium and manganese ions but not by calcium ions. R alpha 2I is recognised by anti-functional anti-alpha 2 monoclonal antibodies 6F1, 5E8 and P1E6 in capture ELISAs, and anti-functional antibodies inhibited r alpha 2I-collagen binding. In addition, r alpha 2I inhibits cell spreading on collagen. R alpha 2I is therefore a collagen-binding domain and can account for many of the collagen-binding functions of integrin alpha 2 beta 1. We have also determined the collagen specificity of r alpha 2I and found that it binds types I, II and XI collagen.

Identification and Characterization of a Human Platelet Glycoprotein VI Peptidomimetic Permitting Molecular Imaging of Fibrosis.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1830-1830
Author(s):  
Martine Jandrot-Perrus ◽  
Julien Muzard ◽  
Laure Sarda-Mantel ◽  
Stéphane Loyau ◽  
Alain Meulemans ◽  
...  
Keyword(s):  
Binding Site ◽  
Cyclic Peptide ◽  
Solid Phase ◽  
Large Field ◽  
Platelet Glycoprotein ◽  
Type I ◽  
Collagen Binding ◽  
Glycoprotein Vi

Abstract Glycoprotein VI (GPVI), the main receptor for platelet activation by collagen, has been shown to play an important role in thrombosis, vascular remodelling and atherothrombosis. GPVI which belongs to the immunoglobulin receptor family, binds to fibrillar type I and type III collagens of vascular as well as non-vascular origin. 9O12.2, a high affinity monoclonal antibody directed to the GPVI extracellular domain, blocks GPVI binding to collagen and possess antithrombotic properties (Ohlmann et al J.Thromb. Haemost. 2008,6:1013). We have hypothesized that the 9O12.2 epitope overlaps, at least in part, with the collagen-binding site on GPVI and (ii) that molecules mimicking the 9O12.2 epitope can be expected to be antithrombotic by competing with platelet GPVI for binding to collagen and/or to act as tracers for collagen in vivo. A bacterial random 12 mer cyclic peptide library was screened against the 9O12.2 IgG. Twenty clones were selected. Sequencing the inserts revealed 9 peptidic motifs with 7 identical residues. One sequence was selected to synthesize a biotin-coupled constrained peptide. (designated collagelin). Surface plasmon resonance (SPR) analysis showed that 9O12.2 IgG bound to immobilized collagelin (KD 10−6M) and that binding was inhibited in the presence of soluble recombinant (sr)GPVI or after disulfide bridge reduction as expected for a molecule mimicking the 9O12.2 epitope known to be conformational (Lecut et al. J.Biol.Chem.2004, 279:52293). Using SPR and solid phase assays, we observed that collagelin bound to immobilized fibrillar collagen (KD10−7M) and that binding was inhibited by 9O12.2 IgG and by rsGPVI, indicating that collagelin mimics at least in part the collagen-binding site of GPVI. Collagelin did not inhibit collagen-induced platelet aggregation in vitro. However, histochemical analysis demonstrated that it bound to collagen on sections of rat aortas and of rat tail tendon. We then hypothesized that collagelin could be retained in vivo at sites of collagen accumulation, thus allowing isotopic imaging of fibrosis. Collagelin and a control peptide (same size and cyclic,) were labeled either indirectly using 99mTc-streptavidin or directly with 99mTc and iv injected into rats presenting fibrotic scars of myocardial infarction. Radiolabeled collagelin uptake in fibrosis areas was demonstrated in vivo by planar and tomographic scintigraphy. Mean heart-to-lung ratios were of 2.76±0.36 and 2.08±0.17 for 99m Tc-streptavidin-coupled collagelin and 99m Tc-collagelin respectively. Ex vivo, autoradiography on frozen heart sections showed a clear uptake of labeled-collagelin in the infarct collagen-rich scars with mean scar to remote myocardium activity ratios of 2.52±0.2 and 2.92±.053 for 99mTc-streptavidin-coupled collagelin and 99mTc-collagelin respectively as compared with 1.82±0.32 and 1.61±0.23 for the control peptides (p<0.006 and 0.01). In conclusion, we have produced a peptide which partly mimics the collagen binding site of GPVI, specifically binds to collagen and appears to be a specific tool for direct targeting of collagen in vitro and in vivo. Collagelin or derived molecules thus potentially have a large field of applications, as a tracer of fibrotic lesions, in non-invasive vascular as vascular pathologies.

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