Identification of fibrinogen derivatives in the Triton-insoluble residue of human blood platelets.
Several proteins (eg. actin, myosin, and actin-binding protein) in the Triton-insoluble residue of thrombin-stimulated platelets are important in the formation of cytoskeletal structures. Electrophoretic analyses have shown that unidentified protein bands of 68,000, 55,000, and 48-50,000 daltons are also present in larger amounts after thrombin stimulation. Since these molecular weights correspond roughly to those of the alpha, beta, and gamma chains of fibrin, and since fibrinogen is found in platelet alpha-granules, these bands were compared to those obtained when purified fibrinogen was treated with thrombin, exposed to 1% Triton X-100-5 mM EGTA, and the resultant Triton-insoluble residue sedimented. Identification of the 68,000-, 55,000-, and 48-50,000-dalton bands as fibrinogen derivatives was confirmed by identifying them in comigration studies and in autoradiographs of Triton-insoluble residues of platelets that were electrophoretically transferred to nitrocellulose paper and treated with antifibrinogen antibody and 125I-protein A. Furthermore, if the platelet suspension was treated with thrombin in the presence of calcium ions, protein bands characteristic of the action of Factor XIII on fibrin were observed, active platelet Factor XIII apparently having been made available by lysis of platelets during preparations. Making use of the electrophoretic properties of tubulin recently described by Best et al [1981], comigration studies using hog brain tubulin indicated that tubulin is not present in significant amounts in the Triton-insoluble residue of platelets as previously suggested. The identification of these proteins as fibrinogen derivatives does not demonstrate a physiological interaction between fibrin and the platelet cytoskeleton, since fibrin is Triton-insoluble and can be pelleted even in the absence of platelet cytoskeletons.
D001792:Blood Platelets / Q000032:analysis*; D003599:Cytoskeleton / Q000032:analysis; D005176:Factor XIII / Q000032:analysis; D005340:Fibrinogen / Q000302:isolation & purification*; D006801:Humans; D008970:Molecular Weight; D017830:Octoxynol; D011092:Polyethylene Glycols; D012995:Solubility; D013917:Thrombin; D014404:Tubulin / Q000097:blood
D014404:Tubulin; D011092:Polyethylene Glycols; D005340:Fibrinogen; D017830:Octoxynol; D005176:Factor XIII; D013917:Thrombin