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Type I and type III collagen‐platelet interaction: Inhibition by chemically synthesized and recombinant hybrid peptide
Author(s) -
Zhu Jiaqian,
Du Haiming,
Chiang Thomas M.
Publication year - 2006
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.20.4.a521-e
Subject(s) - peptide , recombinant dna , microbiology and biotechnology , chemistry , biochemistry , platelet , collagen receptor , receptor , polyclonal antibodies , type i collagen , peptide sequence , biology , antibody , integrin , gene , immunology , endocrinology
We have previously cloned and characterized a platelet receptor for type III collagen (47‐kDa) from a human bone marrow cDNA phage library and defined two active peptides. We also cloned and characterized a platelet receptor for type I collagen (65‐kDa) and defined an active peptide. Our objective was to study whether there is type specificity of these active peptides. We have engineered a mutant receptor clone by replacing the active peptides of the platelet receptor for type III collagen with the active peptide of the platelet receptor for type I collagen at the amino terminal end and expressed a recombinant (rMIII). The rMIII does not alter the hydrophilicity pattern of the protein. The purified recombinant protein reacts with polyclonal anti‐47‐kDa and anti‐65‐kDa active peptide antibodies. The purified recombinant protein inhibits both types I and III collagen‐induced platelet aggregation. This rMIII also inhibits the adhesion of washed platelets to rabbit aortic segments (nature matrix) in a dose‐dependent manner. A hybrid peptide was constructed and expressed in a prokaryotic system. The purified recombinant hybrid peptide contained each active peptide of platelet type I and type III collagen receptors also inhibits types I and III collagen‐induced platelet aggregation as well as 47‐kDa and 22‐kDa phosphorylation. These results suggest that there is a type specific reactive site on platelets for type I and type III collagens. (Supported by grants from VA and NIH‐NIAMS‐SCOR).