Proton-translocating ATPase from bovine kidney medulla: partial purification and reconstitution.
Proton-translocating ATPase from bovine kidney medulla: partial purification and reconstitution.
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来自牛肾髓质的质子转位 ATP 酶：部分纯化和重建。
DOI：
10.1152/ajprenal.1988.254.1.f71
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Caldwell,J
中科院分区：
文献类型：
--
作者：
Gluck,S;Caldwell,J
从牛肾髓质微粒体中部分纯化出负责尿液和空泡酸化的质子易位atp酶。纯化后的atp酶活性最大比活性为1.7 mmol .min-1。并被n -乙基马来酰亚胺完全抑制。高压排粒径液相色谱法测得完整蛋白的相对分子量（Mr）为586,000。分离酶的非变性凝胶在MrS为551,000和523,000时显示出两条蛋白带。分离的H+- atp酶的十二烷基硫酸钠凝胶电泳显示了MrS为70,000,56,000,45,000,42,000,38,000,31,000,15,000,14,000和12,000的组分亚基。分离的H+- atp酶的性质与微粒体依赖atp的质子转运密切相关。分离的H+- atp酶被重组成磷脂脂质体，并证明n -乙基马来酰亚胺抑制atp依赖性电位产生，与电致质子运输一致。在整体结构上，该酶似乎是一种新型的H+- atp酶，具有F0F1类离子易位atp酶的几个特征，但在免疫和结构上与线粒体f1 - atp酶不同。
The proton-translocating ATPase that is responsible both for urinary and vacuolar acidification was partially purified from bovine kidney medulla microsomes. ATPase activity was purified to a maximum specific activity of 1.7 mumol.min-1.mg prot-1 and was inhibited completely by N-ethylmaleimide. The relative molecular weight (Mr) of the intact protein estimated by high-pressure size-exclusion liquid chromatography was 586,000. Nondenaturing gels of the isolated enzyme revealed two protein bands at MrS of 551,000 and 523,000. Sodium dodecyl sulfate-gel electrophoresis of the isolated H+-ATPase revealed component subunits at MrS of 70,000, 56,000, 45,000, 42,000, 38,000, 31,000, 15,000, 14,000, and 12,000. The properties of the isolated H+-ATPase and of microsomal ATP-dependent proton transport correlated closely. The isolated H+-ATPase was reconstituted into phospholipid liposomes and demonstrated N-ethylmaleimide-inhibitable ATP-dependent potential generation, consistent with electrogenic proton transport. In overall structure, the enzyme appears to be a new type of H+-ATPase with several features of the F0F1 class of ion-translocating ATPases but is immunologically and structurally different from the mitochondrial F1-ATPase.