Effects of diamide and dibucaine on platelet glycoprotein Ib, actin-binding protein and cytoskeleton.
During extraction of platelets by 1% Triton X-100, the actin-binding protein (platelet filamin) and a 230 kDa protein are degraded by a calcium-activated thiol protease. Occurrence of degradation products of Mr 190 000 (HF-1) and 90 000 (HF-2) is a sensitive indicator of this proteolysis, and can be used to decide whether reduced amounts of the actin-binding protein in extracts are due to proteolysis or to incorporation in the Triton-insoluble (cytoskeletal) fraction. Diamide, which is a sulfhydryl-oxidizing protein cross-linker, inhibits the calcium-activated protease, polymerizes the actin-binding protein and the 230 kDa protein, increases the incorporation of glycoprotein Ib into the cytoskeletal fraction, and inhibits platelet agglutination induced by bovine von Willebrand factor. Inhibition of platelet agglutination by pretreatment with diamide is partly reversed by dibucaine which activates the calcium-activated protease. These observations are in accordance with a working hypothesis that interactions of glycoprotein Ib with cytoskeleton affect, and possibly regulate, its receptor function in the intact platelet.
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D000818:Animals; D001391:Azo Compounds / Q000494:pharmacology*; D001792:Blood Platelets / Q000032:analysis; D002154:Calpain; D002352:Carrier Proteins / Q000378:metabolism*; D002417:Cattle; D003958:Diamide / Q000494:pharmacology*; D003992:Dibucaine / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000378:metabolism; D018260:Gelsolin; D006023:Glycoproteins / Q000378:metabolism*; D006801:Humans; D007123:Immunoelectrophoresis, Two-Dimensional; D008840:Microfilament Proteins; D008970:Molecular Weight; D017830:Octoxynol; D010974:Platelet Aggregation / Q000187:drug effects; D010980:Platelet Membrane Glycoproteins; D011092:Polyethylene Glycols; D011817:Rabbits; D014841:von Willebrand Factor / Q000494:pharmacology
D001391:Azo Compounds; D002352:Carrier Proteins; D018260:Gelsolin; D006023:Glycoproteins; D008840:Microfilament Proteins; D010980:Platelet Membrane Glycoproteins; C016953:brevin; D014841:von Willebrand Factor; D003958:Diamide; D011092:Polyethylene Glycols; D017830:Octoxynol; D010450:Endopeptidases; D002154:Calpain; D003992:Dibucaine