利用CHAPS分离豌豆叶绿体偶联因子复合物  

Isolation of the CF_0-CF_1 Complex from Pea Chloroplasts by Using A Newly Developed Detergent CHAPS

作　　者：王国强[1] SelmanBruceR. 

机构地区：[1]中国科学院上海植物生理研究所,上海200032 [2]美国威斯康星大学生物化学系

出　　处：《植物生理学报（0257-4829)》1989年第1期93-97,共5页Acta Phytophysiologica Sinica

基　　金：美国NIH基金和中国国家自然科学基金

摘　　要：近年来不少工作者对叶绿体偶联因子复合物的结构、功能和发生的问题颇为关注(程秋琛等 1986,Merchant等 1985,Nelson 1982,Pick等1979,Strontman等1983,Suss等 1983)。The term chloroplast CF_0-CF_1 complex of higher plant is used to designate a complex of energy-transducing ATP Synthase which is associated with a movement of proton across thylakoid membranes. The complex can be functionally divided into two parts: an integral membrane sector (CF_0), which is embedded in the lipid core of the membrane and acts as a proton channel, and an extrinsic membrane sector (CF_1), which emerges from the membrane and contains the catalytic sites responeible for ATP synthesis or hydrolysis. There exists no simple and economical procedure for the isola- tion of a highly active CF_0-CF_1 complex from higher plant with the full subunit composition. In this paper we described a procedure for the isolation and purification of CF_0-CF_1 com- plex from higher plant chloroplasts. The CF_0- CF_1 complex from pea chloroplasts has been isolated by using a newly developed detergent CHAPS which was introduced as an alternative to some detergents such as cholate, Triton X-100 and octylglucoside to solubize membrane pro- tein. The procedure for the isolation of CF_0- CF_1 complex from pea chloroplasts involves four main steps. The first Involves washing chloroplast membranes with 10 mmol/L Na- PPi (pH 8.0) buffer three times. The mem- branes which were washed thoroughly yielded a final CF_0-CF_1 complex preparation with very little high molecular weight contaminants. The second involved using. CHAPS to solubilize the membrane-bound CF_0-CF_1 complex. There is a constant optimum contentration lying be- tween 0.1%～1.0%. The third involved using 37%～45% (NH_4)_2 SO_4 to precipitate the CF_0- CF_1 protein from the CHAPS extract. The protein precipitate by 45% (NH_4)_2SO_4 showe

关 键 词：洗涤剂 叶绿体 豌豆 

分 类 号：Q732[生物学—分子生物学]