Effect of a hybrid peptide derived from platelet type I and type III collagen receptors on CHO-65 cells.
The function of the platelet collagen receptor, 65-kDa, specific for type I collagen (CI), in eukaryotic cells remains elusive. In order to examine the effect of posttranslational modification of the receptor, we have made a construct, which contains the 65-kDa cDNA ligated into pcDNA3 (eukaryotic expression vector) by using standard cloning methodologies. The construct was transfected into CHO (CHO-65) cells, which expressed the protein, detectable by immunoblots [using the anti-65kDa antibodies (65-Ab)], and flow cytometry confirmed expression on the cell surface. A binding assay was used to evaluate the interaction of the expressed protein with CI. A chemically synthesized hybrid peptide (cHyB) containing the active peptides derived from the platelet receptors for types I and III collagen (9 amino acids each with a linker of 10 amino acid residues) inhibited the binding of the 65-Ab and FITC-CI on the CHO-65 cells. These results suggest that the 65-kDa proteins play a role in the interaction with CI and could potentially lead to new therapeutic development for an anti-platelet agent.
Chiang
Thomas M
TM
Department of Veterans Affairs and the Departments of Medicine and Molecular Sciences, University of Tennessee-Health Science center, Memphis, TN 38104, USA. tchiang@utmem.edu
D000818:Animals; D001792:Blood Platelets / Q000378:metabolism*; D016466:CHO Cells; D024042:Collagen Type I / Q000235:genetics / Q000378:metabolism*; D024061:Collagen Type III / Q000235:genetics / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D009154:Mutation; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000235:genetics / Q000502:physiology*; D011485:Protein Binding / Q000235:genetics; D038981:Receptors, Collagen / Q000235:genetics / Q000378:metabolism*
D024042:Collagen Type I; D024061:Collagen Type III; D010446:Peptide Fragments; D038981:Receptors, Collagen