用AMP-琼脂糖亲和层析法从大鼠肝线粒体中纯化一种可溶性ATP酶。

Purification of a soluble ATPase from rat liver mitochondria by AMP-Sepharose affinity chromatography.

作者信息

Le Deaut J Y, Egly J M, Ledig M, Mandel P

出版信息

Biochim Biophys Acta. 1978 Aug 7;525(2):438-45. doi: 10.1016/0005-2744(78)90240-1.

Abstract

ATPase (ATP phosphohydrolase, EC 3.6.1.3) activity was shown in the soluble fraction of rat liver micochondria. Two molecular forms (ATPase 1 and 2) were isolated. ATPase 1 has already been studied. The present paper deals with the purification method of ATPase 2 which was achieved by the following steps: (NH4)2SO4 precipitation. DEAE-cellulose chromatography, hydroxyapatite chromatography, Sephadex G100 filtration and AMP-Sepharose affinity chromatography. The purified protein was characterized by bidimensional polyacrylamide gel electrophoresis. Molecular weight evaluated by SDS-polyacrylamide gel electrophoresis and Sephadex G100 gel filtration was found to be 61 500 +/- 3000.

摘要

ATP酶（ATP磷酸水解酶，EC 3.6.1.3）活性在大鼠肝脏线粒体的可溶性部分中被检测到。分离出了两种分子形式（ATP酶1和ATP酶2）。ATP酶1已经被研究过。本文论述了ATP酶2的纯化方法，该方法通过以下步骤实现：硫酸铵沉淀、二乙氨基乙基纤维素色谱法、羟基磷灰石色谱法、葡聚糖凝胶G100过滤和AMP-琼脂糖亲和色谱法。通过双向聚丙烯酰胺凝胶电泳对纯化后的蛋白质进行了表征。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和葡聚糖凝胶G100凝胶过滤评估的分子量为61500±3000。