Glycoprotein V is not the thrombin activation receptor on human blood platelets.
Thrombin activation of platelets involves two receptors: glycoprotein Ib (GPIb), which affects the kinetics of the response; and, as a strong candidate for the second, essential receptor, GPV, a hydrophobic, 82-kd glycoprotein with an isoelectric point (pI) of pH 5.85 to 6.55. Whole platelets were treated with endogenous platelets calcium-activated proteases, yielding a major fragment, GPV8, with molecular weight (mol wt) of 79 kilodaltons (kd). The fragment was purified by affinity chromatography on wheat germ agglutinin followed by ion exchange chromatography on DEAE-Sephacel using first a 0 to 0.7-mol/L and then a 0 to 0.3-mol/L NaCl gradient. A rabbit was immunized with the purified GPV8 for preparation of polyclonal antibodies. Crossed immunoelectrophoresis and two-dimensional polyacrylamide gel electrophoresis (PAGE) electrophoretic blotting with the separate phases of a Triton X-114 phase partition of human platelets showed the characteristic pattern of GPV in the hydrophobic phase. During thrombin-induced platelet aggregation GPV is hydrolysed, releasing a fragment, GPVf1, to the supernatant. The fragment GPVf1 still contains a thrombin-binding site. Anti-GPV antibodies blocked GPV proteolysis, but did not inhibit platelet activation induced by thrombin. We conclude that proteolysis of GPV by thrombin is not essential for platelet activation.
D001792:Blood Platelets / Q000502:physiology*; D002154:Calpain / Q000378:metabolism; D002462:Cell Membrane / Q000502:physiology; D006023:Glycoproteins / Q000502:physiology*; D006801:Humans; D007158:Immunologic Techniques; D007526:Isoelectric Point; D008565:Membrane Proteins / Q000502:physiology*; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis / Q000276:immunology; D010974:Platelet Aggregation; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface / Q000502:physiology; D013917:Thrombin / Q000502:physiology*
D006023:Glycoproteins; D008565:Membrane Proteins; D010446:Peptide Fragments; D010980:Platelet Membrane Glycoproteins; D011956:Receptors, Cell Surface; D013917:Thrombin; D002154:Calpain