Redistribution of granulophysin and SRC protein in normal and gray platelets after activation

In this study, two mAbs that recognize specifically the src protein pp60(c-src) in a wide variety of cells (mAb 327 and GD11) have been used to vizualize the src protein expression on human platelets by immunogold electron microscopy. The mAb D545 directed against the dense granule membrane p40 protein granulophysin was used as a control. Almost no pp60(c-src) could be detected on the plasma membrane from resting platelets. However, it appeared on the platelet surface after thrombin stimulation and was found preferentially on the pseudopods. The distribution of the src protein on thrombin-activated platelets was similar to that of granulophysin, the dense granule protein, although in a much lesser extent. In platelets from patients with the gray-platelet syndrome, devoid of alpha-granules, pp60(c-src) was absent on resting platelet surface but after thrombin activation expressed at the membrane surface to a normal extent. The results suggest that the src protein is indeed located in an intraplatelet component which is liberated during platelet stimulation, this finding being compatible with its dense granule localization.