The hydrogen ion-pumping adenosine triphosphatase of platelet dense granule membrane. Differences from F1F0- and phosphoenzyme-type ATPases.
Using a coupled transport assay which detects only those ATPase molecules functionally inserted into the platelet dense granule membrane, we have characterized the inhibitor sensitivity, substrate specificity, and divalent cation requirements of the granule H+ pump. Under identical assay conditions, the granule ATPase was insensitive to concentrations of NaN3, oligomycin, and efrapeptin which almost completely inhibit ATP hydrolysis by mitochondrial membranes. The granule ATPase was inhibited by dicyclohexylcarbodiimide but only at concentrations much higher than those needed to maximally inhibit mitochondrial ATPase. Vanadate (VO3-) ion and ouabain also failed to inhibit granule ATPase activity at concentrations which maximally inhibited purified Na+,K+-ATPase. Two alkylating agents, 7-chloro-4-nitrobenz-2-oxa-1,3-diazole and N-ethylmaleimide both completely inhibited H+ pumping by the granule ATPase under conditions where ATP hydrolysis by mitochondrial membranes or Na+,K+-ATPase was hardly affected. These results suggest that the H+-pumping ATPase of platelet granule membrane may belong to a class of ion-translocating ATPases distinct from both the phosphoenzyme-type ATPases present in plasma membrane and the F1F0-ATPases of energy-transducing membranes.
D000251:Adenosine Triphosphatases / Q000097:blood; D000818:Animals; D001792:Blood Platelets / Q000201:enzymology* / Q000648:ultrastructure; D002413:Cations, Divalent / Q000378:metabolism; D003594:Cytoplasmic Granules / Q000201:enzymology; D008930:Mitochondria, Liver / Q000201:enzymology; D009711:Nucleotides / Q000378:metabolism; D006180:Proton-Translocating ATPases / Q000097:blood*; D012701:Serotonin / Q000097:blood; D000254:Sodium-Potassium-Exchanging ATPase / Q000097:blood; D013552:Swine
D002413:Cations, Divalent; D009711:Nucleotides; D012701:Serotonin; D000251:Adenosine Triphosphatases; D006180:Proton-Translocating ATPases; D000254:Sodium-Potassium-Exchanging ATPase