Identification of glycoprotein IV (CD36) as a primary receptor for platelet-collagen adhesion.
The role of glycoprotein IV (GPIV) in platelet activation processes has been examined by several different approaches: (i) Fab fragments of a monospecific polyclonal antibody to purified platelet GPIV (approximately 20 micrograms/ml) completely inhibited platelet shape change, aggregation, and secretion induced by collagen. Aggregation and secretion by ADP (but not shape change) and by epinephrine were also inhibited, but there was no effect on platelet activation induced by thrombin, arachidonate, or ionophore A23187. (ii) Purified GPIV was able to compete completely with membrane-bound GPIV to inhibit platelet activation induced by collagen, including shape change, but not in activation induced by any of the other platelet agonists. 50% inhibition of collagen-induced activation and secretion were obtained at GPIV concentrations of approximately 10 nM (1 micrograms/ml). (iii) Purified GPIV bound rapidly and reversibly to collagen Type I fibrils, and binding was not inhibited by adhesive proteins such as denatured collagen, fibronectin, fibrinogen, or von Willebrand factor. The direct binding of purified GPIV to collagen Type I fibrils fit best to a single site model with Kd 0.34 +/- 0.10 nM. (iv) Using a microtiter assay, platelet adhesion to collagen was shown to be inhibited by Fab fragments of monospecific polyclonal anti-GPIV antibodies, but adhesion to other adhesive proteins was unaffected. (v) When anti-GPIV was added at various times during adhesion the time dependence of inhibition was seen to be biphasic. Anti-GP antibody was able to reverse adhesion that occurred within the first 5-8 min and to inhibit adhesion occurring thereafter. These results demonstrate that GPIV mediates the early stages of platelet recognition by and attachment to collagen but that there may be a second GPIV-independent mechanism that mediates the subsequent anchorage of these adherent platelets.
Tandon
N N
NN
Cell Biology Laboratory, American Red Cross, Rockville, Maryland 20855.
Kralisz
U
U
Jamieson
G A
GA
D000937:Antigen-Antibody Reactions; D000943:Antigens, Differentiation / Q000502:physiology*; D000954:Antigens, Surface / Q000502:physiology*; D001792:Blood Platelets / Q000502:physiology*; D018955:CD36 Antigens; D015815:Cell Adhesion Molecules; D003094:Collagen / Q000378:metabolism*; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000494:pharmacology; D066298:In Vitro Techniques; D010973:Platelet Adhesiveness; D010980:Platelet Membrane Glycoproteins / Q000502:physiology*; D013997:Time Factors
D000943:Antigens, Differentiation; D000954:Antigens, Surface; D018955:CD36 Antigens; D015815:Cell Adhesion Molecules; D007140:Immunoglobulin Fab Fragments; D010980:Platelet Membrane Glycoproteins; D003094:Collagen