Identification of talin as a major cytoplasmic protein implicated in platelet activation.
During platelet activation there is a major reorganization in the platelet cytoskeleton that accompanies a rapid change in platelet shape. Many of the events associated with activation are attributed to a rise in calcium concentration within the platelet cytoplasm. One direct consequence of the elevated calcium is the activation of a calcium-dependent protease that cleaves a major platelet protein of relative molecular mass (Mr) approximately 235,000 (235K) to 200K. This protein, P235, has been purified and reported to interact with actin, but the significance of the proteolytic cleavage is unknown. Talin, a cytoskeletal protein in smooth muscle and fibroblasts, binds vinculin and, together with vinculin, is localized in fibroblasts at sites of actin-membrane attachment. Talin and P235 have similar purification procedures, sedimentation coefficients and Stokes' radii (ref. 6 and Molony et al., unpublished observations). Of particular significance, talin is readily cleaved by proteases from approximately 215K to a fragment of approximately 190K. Given these similarities we have investigated the possible relationship between these proteins. Here we demonstrate that platelet P235 is recognized by anti-talin antibody and that it binds vinculin. Both proteins are cleaved in vitro by the calcium-activated protease to yield similar fragments. We conclude that P235 corresponds to the platelet form of talin.
D000199:Actins / Q000378:metabolism; D001792:Blood Platelets / Q000166:cytology* / Q000187:drug effects; D002118:Calcium / Q000378:metabolism; D002154:Calpain / Q000378:metabolism; D003285:Contractile Proteins / Q000032:analysis; D003429:Cross Reactions; D003593:Cytoplasm / Q000378:metabolism; D003599:Cytoskeleton / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D008970:Molecular Weight; D009124:Muscle Proteins / Q000378:metabolism / Q000494:pharmacology*; D016608:Talin; D016596:Vinculin
D000199:Actins; D003285:Contractile Proteins; D009124:Muscle Proteins; D016608:Talin; D016596:Vinculin; D002154:Calpain; D002118:Calcium