Platelet Shape Change Induced by the Peptide YFLLRNP

Abstract

Platelet shape change represents an early response to activating agents. Whereas the PAR-1-activating peptide SFLLRN induces a total platelet activation, YFLLRNP brings the process only to the shape change step in a process independent of the cytosolic Ca 2+ concentration. In this paper, the YFLLRNP-induced shape change has been observed in human citrated platelet-rich plasma (PRP). Scanning electron microscopy of platelets activated by 300 μM YFLLRNP showed platelets that had changed shape and extended long pseudopods. The protein content of the material sedimenting at 13,000× g from 1% Triton X-100 extracts increased during the shape change, indicating a reorganization of the cytoskeleton. This was supported by electrophoresis. The GP IIb–IIIa complex was not activated, however, and the platelets that had undergone shape change did not support clot retraction. As no increase in binding of FITC-labeled annexin V (FITC-annexin V) was observed, the extensive shape change was not associated with a disturbance of the membrane phospholipid asymmetry. Platelet aggregation was never observed with 300 μM YFLLRNP, but could be seen at much higher concentrations, neither was secretion from dense granules observed at 300 μM as no extracellular ATP could be observed. These studies confirm and extend the concept of the Ca 2+-independent shape change as a distinct and sharply delineated process that, in itself, may be of little pathophysiological importance if such “partly activated” platelets occur in the circulation.