Complement C5b-9-stimulated platelet secretion is associated with a Ca2+-initiated activation of cellular protein kinases.
Membrane assembly of the C5b-9 proteins on gel-filtered human platelets has been shown to initiate the nonlytic release of alpha-granule contents and expression of membrane prothrombinase sites, suggesting cellular activation by these ostensibly cytolytic plasma proteins (Wiedmer, T., Esmon, C. T., and Sims, P. J. (1986) J. Biol. Chem. 261, 14587-14592). We now examine the mechanism of the C5b-9-induced release reaction. The release of alpha-granule contents upon C5b-9 assembly is accompanied by expression of alpha-granule membrane glycoprotein 140 on the platelet surface, confirming that the complement-mediated release reaction occurs by secretory fusion of the alpha-granule with the plasma membrane. C5b-9 binding initiates the phosphorylation of both 40- and 20-kDa platelet proteins, indicative of activation of protein kinase C and myosin light chain kinase, respectively. Activation of cellular protein kinases under these conditions was not accompanied by the formation of inositol phosphates and was found to strictly depend upon extracellular Ca2+, suggesting that the platelet's secretory response to the C5b-9 proteins is triggered directly by the influx of Ca2+ across the plasma membrane. measurement of intracellular Ca2+ confirmed that elevation of this ion in the cytosol was strictly dependent upon increased plasma membrane permeability due to C5b-9 assembly and was not accompanied by mobilization of this ion from internal storage pools. The C5b-9-mediated secretory response was blocked by sphingosine, a potent inhibitor of protein kinase C, but was unaffected by the cyclooxygenase inhibitor indomethacin, suggesting that feedback (receptor-linked) by thromboxane is not required for platelet activation after C5b-9 insertion.
D000328:Adult; D001792:Blood Platelets / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D001798:Blood Proteins / Q000378:metabolism; D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000494:pharmacology*; D015938:Complement Membrane Attack Complex; D003165:Complement System Proteins / Q000502:physiology*; D003594:Cytoplasmic Granules / Q000378:metabolism; D004789:Enzyme Activation; D005456:Fluorescent Dyes; D006801:Humans; D007211:Indoles; D007295:Inositol Phosphates / Q000097:blood; D007700:Kinetics; D010766:Phosphorylation; D011494:Protein Kinases / Q000097:blood*; D013050:Spectrometry, Fluorescence; D013917:Thrombin / Q000502:physiology
D001798:Blood Proteins; D015938:Complement Membrane Attack Complex; D005456:Fluorescent Dyes; D007211:Indoles; D007295:Inositol Phosphates; D000001:Calcimycin; D003165:Complement System Proteins; D011494:Protein Kinases; D013917:Thrombin; C048960:indo-1; D002118:Calcium