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编码寡霉素敏感相关蛋白(OSCP)的大鼠cDNA在酿酒酵母中的功能表达。

The functional expression of a rat cDNA encoding OSCP in the yeast Saccharomyces cerevisiae.

作者信息

Prescott M, Higuti T, Nagley P, Devenish R J

机构信息

Department of Biochemistry and Molecular Biology, Monash University, Clayton, Victoria, Australia.

出版信息

Biochem Biophys Res Commun. 1995 Feb 27;207(3):943-9. doi: 10.1006/bbrc.1995.1276.

Abstract

The oligomycin sensitivity conferring protein (OSCP) of Saccharomyces cerevisiae is a component of the stalk FA sector of the mitochondrial F1Fo-ATP synthase complex (mtATPase). The subunits comprising this sector have been implicated in the coupling of proton flux through the membrane Fo sector to ATP synthesis on the catalytic F1 sector. We tested whether the yeast subunit encoded by the ATP5 gene could be functionally replaced by a mammalian homologue, rat OSCP. The yeast and rat OSCP proteins have 58% homology in terms of conserved amino acids. A vector-borne rat cDNA encoding the OSCP precursor was introduced into a yeast strain lacking endogenous OSCP as a result of its disrupted ATP5 gene. The resultant yeast cells showed essentially a wildtype growth rate at 18, 28 and 35 degrees C, on nonfermentable substrates. Expression of mammalian FA subunit homologues in yeast should provide a productive new approach to generating information on the structure and function of mtATPase.

摘要

酿酒酵母的寡霉素敏感性赋予蛋白(OSCP)是线粒体F1Fo - ATP合酶复合体(mtATPase)柄部FA区域的一个组成部分。构成该区域的亚基与质子通过膜部Fo区域的通量与催化性F1区域上的ATP合成之间的偶联有关。我们测试了由ATP5基因编码的酵母亚基是否能被哺乳动物同源物大鼠OSCP在功能上替代。酵母和大鼠的OSCP蛋白在保守氨基酸方面具有58%的同源性。将编码OSCP前体的载体携带的大鼠cDNA导入因ATP5基因破坏而缺乏内源性OSCP的酵母菌株中。所得酵母细胞在18、28和35摄氏度下,在非发酵底物上显示出基本野生型的生长速率。在酵母中表达哺乳动物FA亚基同源物应该为生成有关mtATPase结构和功能的信息提供一种有效的新方法。

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