Open AccessAssembly of cytochrome‐ c oxidase in cultured human cells
Author(s) -
Nijtmans Leo G. J.,
Taanman JanWillem,
Muijsers Anton O.,
Speijer Dave,
Van den Bogert Coby
Publication year - 1998
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1046/j.1432-1327.1998.2540389.x
Subject(s) - cytochrome c oxidase , protein subunit , oxidase test , biochemistry , microbiology and biotechnology , cytochrome b , cytochrome c , cytochrome , polyacrylamide gel electrophoresis , mitochondrion , chemistry , electron transport complex iv , biology , cytosol , enzyme , mitochondrial dna , gene
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.