Some properties of membrane‐bound, solubilized and reconstituted into liposomes H+‐ATPase of vacuoles of Saccharomyces carlsbergensis
Some properties of membrane‐bound, solubilized and reconstituted into liposomes H+‐ATPase of vacuoles of Saccharomyces carlsbergensis
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卡尔斯伯酵母菌液泡膜结合、溶解和重组脂质体 H+-ATP 酶的一些特性
DOI：
10.1016/0014-5793(84)81164-3
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发表时间：
1984
期刊：
影响因子：
3.5
通讯作者：
L. Okorokov
中科院分区：
文献类型：
--
作者：
L. Lichko;L. Okorokov
酵母液泡液泡具有较高的ATPase活性(最高可达1minmol.mg蛋白-1.μ−1)。膜结合和溶解ATPase活性对钒和叠氮不敏感，但被NO3-−、K+和环磷酸腺苷抑制。Dio-9使液泡ATPase的膜形式激活1.5-2倍，但不影响溶解的酶。以大豆磷脂为原料，通过冻融法重组液泡型ATPase。Dio-9、原载体FCCP和离子载体valinmycin和nigericin能有效地刺激天然液泡和蛋白脂质体中的ATPase活性。ACMA荧光猝灭也显示依赖于ATP的H+转运到蛋白脂质体中。液泡型和部分纯化的ATPase制剂也具有GTPase活性。然而，与ATPase不同的是，GTPase不是作为质子泵掺入脂质体中的。
Vacuoles of yeast grown in peptone medium possessed high ATPase activity (up to 1 μmol·mg protein‐1·min−1). Membrane‐bound and solubilized ATPase activities were insensitive to vanadate and azide, but were inhibited by NO3−, K+and cyclic AMP stimulated both membrane‐bound and solubilized ATPase activities. Dio‐9 activated the membrane form of vacuolar ATPase 1.5‐2‐fold and did not affect the solubilized enzyme. Solubilized and partially purified vacuolar ATPase was reconstituted with soy‐bean phospholipids by a freeze‐thaw procedure. ATPase activities in native vacuoles and proteoliposomes were stimulated effectively by Dio‐9, the protonophore FCCP and ionophores valinomycin and nigericin. ATP‐dependent H+transport into proteoliposomes was also shown by quenching of ACMA fluorescence. Vacuolar and partially purified ATPase preparations possessed also GTPase activity. Unlike ATPase, however, GTPase was not incorporated as a proton pump into liposomes.