Assembly of cytochrome-c oxidase in cultured human cells

被引:208
作者
Nijtmans, LGJ
Taanman, JW
Muijsers, AO
Speijer, D
Van den Bogert, C
机构
[1] Univ Amsterdam, Acad Med Ctr, Dept Neurol, NL-1105 AZ Amsterdam, Netherlands
[2] Univ Amsterdam, Acad Med Ctr, Dept Biochem, NL-1105 AZ Amsterdam, Netherlands
[3] Royal Free Hosp, Sch Med, Dept Clin Neurosci, London, England
[4] BioCtr Amsterdam, EC Slater Inst, Amsterdam, Netherlands
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1998年 / 254卷 / 02期
基金
英国惠康基金;
关键词
cytochrome-c oxidase; assembly; respiratory chain; cultured human cell;
D O I
10.1046/j.1432-1327.1998.2540389.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The assembly of cytochrome-c oxidase was studied in human cells cultured in the presence of inhibitors of mitochondrial or cytosolic protein synthesis. Mitochondrial fractions were resolved using two-dimensional PAGE (blue native PAGE and tricine/SDS/PAGE) and subsequent western blots were developed with monoclonal antibodies against specific subunits of cytochrome-c oxidase. Proteins were also visualized using metabolic labeling followed by two-dimensional electrophoresis and fluorography. These techniques allowed identification of two assembly intermediates of cytochrome-c oxidase. Assembly of the 13 subunits of cytochrome-c oxidase starts with the association of subunit I with subunit IV. Then a larger subcomplex is formed, lacking only subunits VIa and either VIIa or VIIb.
引用
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页码:389 / 394
页数:6
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