title: Vanadate-sensitive ATPase from chromaffin granule membranes formed a phosphoenzyme intermediate and was activated by phosphatidylserine.
abstract: Vanadate-sensitive ATPase (115 kDa molecular weight) in adrenal chromaffin granules is an intrinsic membrane enzyme with its catalytic site located at the outer surface of the granules. Upon incubation with [gamma-32P]ATP, the purified ATPase formed an alkaline-labile phosphoenzyme intermediate, which was inhibited by vanadate but not by Na+ or K+. Ratio of ATPase or phosphatase activity and formation of phosphoenzyme intermediate was constant during purification after the first glycerol density gradient centrifugation. Phosphatidylserine specifically activated the enzyme about three-fold by increasing the Vmax value without changing the Km for ATP. Other phospholipids, including phosphatidylglycerol, phosphatidylcholine, phosphatidylinositol, and phosphatidylethanolamine, as well as lysophospholipids and detergents, had no effect. These results indicated that the vanadate-sensitive ATPase belongs to the P-type ATPases, which differ from known cation-translocating P-type ATPases.
authors.affiliation: Department of Organic Chemistry and Biochemistry, Osaka University, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Moriyama
authors.forename: N
authors.initials: N
authors.lastname: Nelson
authors.forename: M
authors.initials: M
authors.lastname: Maeda
authors.forename: M
authors.initials: M
authors.lastname: Futai
mesh_terms: D000251:Adenosine Triphosphatases / Q000302:isolation & purification / Q000378:metabolism*; D000313:Adrenal Medulla / Q000201:enzymology; D000818:Animals; D002417:Cattle; D002837:Chromaffin Granules / Q000201:enzymology*; D004789:Enzyme Activation; D007425:Intracellular Membranes / Q000201:enzymology*; D007700:Kinetics; D008970:Molecular Weight; D010718:Phosphatidylserines / Q000494:pharmacology*; D010743:Phospholipids / Q000494:pharmacology*; D010766:Phosphorylation; D013329:Structure-Activity Relationship
chemical_list: D010718:Phosphatidylserines; D010743:Phospholipids; D000251:Adenosine Triphosphatases; C070990:vanadate-sensitive ATPase