Document Detail


Cloning, characterization, and functional studies of a nonintegrin platelet receptor for type I collagen.
MedLine Citation:
PMID:  9239397     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A cDNA (1.6 kb) encoding a platelet protein receptor that binds type I collagen has been isolated from a human bone marrow cDNA library by using a degenerate oligonucleotide probe derived from the amino acid sequence of a CNBr fragment of the purified receptor. Computer search revealed that this cDNA represents the coding sequence of a unique protein. Using the prokaryotic expression system pKK 223-3-65 cDNA, a 54-kD recombinant protein was obtained and purified to apparent homogeneity. In an eukaryotic expression vector (pcDNA3-65 cDNA), a 65-kD protein was identified that was recognized by monoclonal anti-65 kD antibody (anti-65m). The recombinant protein binds to type I, but not to type III collagen by affinity column chromatography. The binding of the recombinant protein to type I collagen-coated Petri dishes is inhibited by anti-65m in a dose-dependent manner. The pcDNA3-65 cDNA-transfected nonadherent T cells express the protein, allowing them to attach to a type I collagen matrix, and are inhibited by anti-65m in a dose-dependent manner. Like the receptor protein purified from platelet membranes, the recombinant protein inhibits type I collagen-induced platelet aggregation and the adhesion of [14C]serotonin-labeled platelets to type I collagen in a dose-dependent manner. The recombinant protein neither binds to type III collagen-coated Petri dishes nor inhibits type III collagen and ADP-induced platelet aggregation, indicating specificity for type I collagen.
Authors:
T M Chiang; A Rinaldy; A H Kang
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Publication Detail:
Type:  Journal Article; Research Support, U.S. Gov't, Non-P.H.S.    
Journal Detail:
Title:  The Journal of clinical investigation     Volume:  100     ISSN:  0021-9738     ISO Abbreviation:  J. Clin. Invest.     Publication Date:  1997 Aug 
Date Detail:
Created Date:  1997-09-04     Completed Date:  1997-09-04     Revised Date:  2009-11-18    
Medline Journal Info:
Nlm Unique ID:  7802877     Medline TA:  J Clin Invest     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  514-21     Citation Subset:  AIM; IM    
Affiliation:
Department of Medicine, University of Tennessee at Memphis, Memphis, Tennessee 38104, USA. tchiang@utmem2.utmen.edu
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MeSH Terms
Descriptor/Qualifier:
Amino Acid Sequence
Base Sequence
Blood Platelets / metabolism*
Cloning, Molecular
Collagen / metabolism*
Humans
Integrins / genetics*,  metabolism
Molecular Sequence Data
Receptors, Cell Surface / genetics*,  metabolism
Receptors, Collagen
Sequence Analysis
Chemical
Reg. No./Substance:
0/Integrins; 0/Receptors, Cell Surface; 0/Receptors, Collagen; 9007-34-5/Collagen
Comments/Corrections

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