PURIFICATION OF THE YEAST PLASMA-MEMBRANE ATPASE SOLUBILIZED WITH A NOVEL ZWITTERIONIC DETERGENT
PURIFICATION OF THE YEAST PLASMA-MEMBRANE ATPASE SOLUBILIZED WITH A NOVEL ZWITTERIONIC DETERGENT
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DOI：
10.1016/0014-5793(80)80763-0
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发表时间：
1980-01-01
期刊：
影响因子：
3.5
通讯作者：
SERRANO, R
中科院分区：
文献类型：
--
作者：
MALPARTIDA, F;SERRANO, R
在粗糙脉孢菌[1，2]、粟酒裂殖酵母[3]和酿酒酵母[4]的质膜中已经鉴定出具有类似动力学性质的ATP酶。在这些真菌细胞中，营养物质的主动运输与质子梯度耦合，因此已经提出该ATP酶作为质子泵起作用[5-71]。为了获得这种重要生理作用的直接证据，有必要纯化酶，将其掺入脂质体中，并在这些结构中寻找ATP驱动的质子转运[8]。在大多数情况下，膜酶的纯化需要用去污剂溶解它们，并且如[9]中所述，必须凭经验找到特定膜蛋白的最佳去污剂。粟酒裂殖酵母的质膜ATP酶已用溶血素[lo]增溶，但该试剂。以及许多其它常规的洗涤剂被用于溶解来自酿酒酵母的酶。
ATPases with similar kinetic properties have been identified in the plasma membranes of Neurospora crassa [1, 2], Schizosaccharomyces pombe [3] andSaccharomyces cerevisiae [4]. In these fungal cells the active transport of nutrients is coupled to the proton gradient and therefore it has been suggested that this ATPase operates as a proton pump [5-71. In order to obtain direct evidence for this important physiological role it would be necessary to purify the enzyme, incorporate it into liposomes and look for ATP-driven proton transport in these structures [8]. The purification of membrane enzymes requires in most cases their solubilization with detergents and, as stated in [9], the optimal detergent for a particular membrane protein has to be found empirically. The plasma membrane ATPase of SchizosaccharomJlces pombe has been solubilized with lysolecithin [lo] but this agent. as well as many other conventional detergents were inoperative in our hands for the solubilization of the enzyme from Saccharomyces cerevisiae.