Assembly of cytochrome-c oxidase in cultured human cells.
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.
Nijtmans
L G
LG
Department of Neurology, Academic Medical Center, University of Amsterdam, The Netherlands. l.g.j.nijtmans@dundee.ac.uk
Taanman
J W
JW
Muijsers
A O
AO
Speijer
D
D
Van den Bogert
C
C
D000911:Antibodies, Monoclonal; D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D003600:Cytosol / Q000201:enzymology; D004318:Doxycycline / Q000494:pharmacology; D003576:Electron Transport Complex IV / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional; D004795:Enzyme Stability; D006801:Humans; D007700:Kinetics; D008928:Mitochondria / Q000201:enzymology; D008954:Models, Biological; D011487:Protein Conformation; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D011500:Protein Synthesis Inhibitors; D003513:Cycloheximide; D003576:Electron Transport Complex IV; D004318:Doxycycline