Fresneau C, Rivière M E, Arrio B
Laboratoire de Bioénergétique Membranaire (URA 1116 du CNRS), Université de Paris-Sud, Orsay, France.
Arch Biochem Biophys. 1993 Oct;306(1):254-60. doi: 10.1006/abbi.1993.1508.
Biochemical properties of the ATPase from the plasma membrane of the cyanobacteria Synechococcus PCC 6311 and PCC 7942 were examined. ATPase activity associated with purified plasma membrane vesicles was strongly inhibited by 100 microM vanadate (87%), 100 microM diethylstilbestrol (70%) and 100 mM fluoride ions (83%). No inhibition was observed in the presence of dicyclohexylcarbodiimide, nitrate, azide, or molybdate. A 50% activation was observed in the presence of 50 mM KCl but none was observed in the presence of NaCl or NH4Cl. This ATPase was able to form a pH gradient, the amplitude of which was decreased by the presence of 100 microM vanadate. On Western blot of the plasmalemma proteins, no labeling was observed with a monoclonal antibody against the beta subunit of the F0-F1 ATPase, although staining was observed with the 55-kDa subunit of the thylakoid membrane ATPase. After phosphorylation of plasmalemma vesicles, by [gamma-32P]ATP, the autoradiograms of the electrophoreses, performed under acid conditions, exhibited labeling of a 110-kDa protein. The results indicated that the Synechococcus plasma membrane ATPase can be classified as a H+ translocating P-type ATPase and compared to the plant plasmalemma ATPase.
对蓝藻聚球藻PCC 6311和PCC 7942质膜上ATP酶的生化特性进行了研究。与纯化的质膜囊泡相关的ATP酶活性受到100微摩尔钒酸盐（87%）、100微摩尔己烯雌酚（70%）和100毫摩尔氟离子（83%）的强烈抑制。在二环己基碳二亚胺、硝酸盐、叠氮化物或钼酸盐存在的情况下未观察到抑制作用。在50毫摩尔氯化钾存在的情况下观察到50%的激活，但在氯化钠或氯化铵存在的情况下未观察到激活。这种ATP酶能够形成pH梯度，100微摩尔钒酸盐的存在会降低其幅度。在质膜蛋白的蛋白质免疫印迹中，未观察到针对F0-F1 ATP酶β亚基的单克隆抗体的标记，尽管观察到类囊体膜ATP酶55千道尔顿亚基的染色。在用[γ-32P]ATP对质膜囊泡进行磷酸化后，在酸性条件下进行的电泳放射自显影片显示出110千道尔顿蛋白质的标记。结果表明，聚球藻质膜ATP酶可归类为一种H+转运P型ATP酶，并可与植物质膜ATP酶进行比较。
