title: Purification and partial characterization of adenosine diphosphatase activity in bovine aorta microsomes.
abstract: Anti-aggregatory activities in bovine aorta microsomal fractions were solubilized with Triton X-100 and separated into two fractions by DEAE-Sepharose CL-6B. One fraction strongly inhibited arachidonic acid-induced platelet aggregation, and the other inhibited ADP-induced aggregation. The latter fraction contained ADPase activity. The ADPase activity was further purified by affinity chromatography. The purified enzyme had specific activities of 43.8 and 48.2 mumol of Pi/min/mg protein for ADP and ATP, respectively. The enzyme required calcium or magnesium ions and it was insensitive to ATPase inhibitors, namely oligomycin and ouabain, and to adenylate kinase inhibitor, Ap5A. Polyacrylamide gel electrophoretic experiments indicated that only one enzyme was involved. This was confirmed by the parallel behavior of ADPase and ATPase activities throughout all the purification steps. These results suggest that the main anti-aggregatory activity of bovine aorta microsomes for ADP-induced aggregation is due to an ATP diphosphohydrolase (EC 3.6.1.5).
authors.forename: Y
authors.initials: Y
authors.lastname: Miura
authors.forename: K
authors.initials: K
authors.lastname: Hirota
authors.forename: Y
authors.initials: Y
authors.lastname: Arai
authors.forename: K
authors.initials: K
authors.lastname: Yagi
mesh_terms: D000244:Adenosine Diphosphate / Q000494:pharmacology; D000818:Animals; D001011:Aorta / Q000166:cytology / Q000201:enzymology*; D001081:Apyrase / Q000302:isolation & purification* / Q000502:physiology; D002118:Calcium / Q000494:pharmacology; D002417:Cattle; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D008274:Magnesium / Q000494:pharmacology; D008861:Microsomes / Q000201:enzymology*; D017830:Octoxynol; D010744:Phosphoric Monoester Hydrolases / Q000302:isolation & purification*; D010974:Platelet Aggregation / Q000187:drug effects; D011092:Polyethylene Glycols / Q000494:pharmacology; D013379:Substrate Specificity
chemical_list: D011092:Polyethylene Glycols; D000244:Adenosine Diphosphate; D017830:Octoxynol; D010744:Phosphoric Monoester Hydrolases; D001081:Apyrase; D008274:Magnesium; D002118:Calcium