The alpha 2 beta 1 integrin cell surface collagen receptor binds to the alpha 1 (I)-CB3 peptide of collagen.
We have previously shown that platelets adhere to collagen substrates via a Mg2(+)-dependent mechanism mediated by the surface glycoprotein Ia-IIa (human leukocyte very late activation protein 2, alpha 2 beta 1 integrin) complex. The adhesion is specific for collagen and is supported by collagen types I, II, III, IV, and VI. Several other members of the integrin family of adhesive protein receptors recognize discrete linear amino acid sequences within their adhesive glycoprotein ligands. Experiments with both intact platelets and with liposomes containing the purified receptor complex indicated that the alpha 2 beta 1 receptor recognized denatured type I collagen in a Mg2(+)-dependent manner. To further localize the binding site, the alpha 1 and alpha 2 chains of type I collagen were purified by gel filtration and ion exchange chromatography and tested as adhesive substrates. Both the alpha 1(I) and alpha 2(I) chains effectively supported Mg2(+)-dependent platelet adhesion. The purified alpha 1(I) collagen chain was then subjected to cleavage with cyanogen bromide, and the resultant peptides were separated by chromatography on carboxymethylcellulose. Only the alpha 1(I)-CB3 fragment supported Mg2(+)-dependent platelet adhesion. The monoclonal antibody P1H5 which recognizes an epitope on the alpha 2 subunit of the integrin receptor and which inhibits the adhesion of both intact platelets and liposomes bearing the purified receptor to collagen also inhibited platelet adhesion to the alpha 1(I)-CB3 fragment. These results indicate that the alpha 2 beta 1 receptor recognizes a sequence of amino acids present in the alpha 1(I)-CB3 fragment of type I collagen. An identical or similar sequence likely mediates binding of the receptor to other collagen polypeptides.
Staatz
W D
WD
Department of Pathology, Washington University School of Medicine, St. Louis, Missouri 63110.
Walsh
J J
JJ
Pexton
T
T
Santoro
S A
SA
D000911:Antibodies, Monoclonal; D001792:Blood Platelets / Q000502:physiology*; D003094:Collagen / Q000378:metabolism*; D006801:Humans; D016023:Integrins / Q000302:isolation & purification / Q000378:metabolism*; D008081:Liposomes; D046911:Macromolecular Substances; D008274:Magnesium / Q000494:pharmacology; D010446:Peptide Fragments / Q000378:metabolism; D010973:Platelet Adhesiveness / Q000187:drug effects; D011956:Receptors, Cell Surface / Q000378:metabolism*; D038981:Receptors, Collagen
D000911:Antibodies, Monoclonal; D016023:Integrins; D008081:Liposomes; D046911:Macromolecular Substances; D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D038981:Receptors, Collagen; D003094:Collagen; D008274:Magnesium