Calcium-Dependent Cross-Linking Processes in Platelets

When platelet cytoplasmic Ca2+ is increased by the ionophore A 23187, there is the coincident appearance of a cross-linked polymer and the partial disappearance of five high molecular weight polypeptide bands (> 145,000). The glycoproteins show a partial disappearance of bands lb, IIb and IV and the total disappearance of hands la and Ilia. The disappearance of the protein bands, possibly contributing to the polymer formation, is prevented by histamine, aminoacetonltrile and cystamlne, which, as pseudodonor amines are known Inhibitors of factor XHIa-catalyzed cross-linking. 14C-histamine, at a tracer concentration, was incorporated into the polymer as well as into myosin, glycoproteins IIb and IIIa (α-actinln), actin and two unidentified low-molecular weight proteins. The polymer formed is also apparent in isolated membranes following the iono-phore-stimulated increase in intracellular Ca2+. These findings are unrelated to a proteolytic activity since the platelet Ca2+-dependent proteases are inhibited by leupep-tin. Ca2+-activation of a platelet cytosol transamidase would explain the data obtained. This platelet transamidase(s) may couple membrane proteins to cytoplasmic contra-tlle proteins. Thus, a new concept is proposed for the stabilization of platelet membranes and platelets as they form the hemostatic plug.