Site of synthesis of the mitochondrial cytochromes in hepatocyte cultures.
The biosynthesis of mammalian mitochondrial cytochromes was explored in primary hepatocyte cultures. When these were pulsed with [35S]methionine in the presence of cycloheximide, eight discrete mitochondrial polypeptides were detected by fluorography after their resolution under denaturing conditions by polyacrylamide gel electrophoresis. Since the pulse labeling of the polypeptides was sensitive to chloramphenicol, an inhibitor of mitochondrial translation, they must be translated on mitochondrial ribosomes. Three were identified as the largest subunits of cytochrome oxidase by their immunoprecipitation with antibody directed against purified rat liver cytochrome oxidase. Another (Mr = 28,000) was identified as one of eight subunits of purified rat liver cytochrome b-c1 complex by its immunoprecipitation with antibody directed against bovine heart b-c1 complex. Since cytochrome b apoprotein is the only product of the mitochondrial genome in the yeast cytochrome b-c1 complex (Krieke, J., Bechmann, H., van Hemert, F. J., Schweyan, R. J., Boer, P. H., Kaudewitz, F., and Groot, G. S. P. (1979) Eur. J. Bio-chem. 101, 607-617), the results strongly suggest that the Mr = 28,000 subunit of liver b-c1 complex is cytochrome b apoprotein. Thus the contribution of the mitochondrial translation system to the cytochrome complexes in liver is identical to that of yeast and Neurospora, and there appears to be no deletion or transfer to the nuclear genome of structural genes for mitochondrially synthesized cytochromes during eukaryotic evolution.
D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D002701:Chloramphenicol / Q000494:pharmacology; D003513:Cycloheximide / Q000494:pharmacology; D003574:Cytochrome c Group / Q000096:biosynthesis; D014450:Electron Transport Complex III; D003576:Electron Transport Complex IV / Q000096:biosynthesis* / Q000302:isolation & purification; D007118:Immunoassay; D008099:Liver / Q000201:enzymology*; D046911:Macromolecular Substances; D008929:Mitochondria, Heart / Q000378:metabolism; D008930:Mitochondria, Liver / Q000201:enzymology*; D009097:Multienzyme Complexes / Q000096:biosynthesis*; D009247:NADH, NADPH Oxidoreductases / Q000096:biosynthesis*; D014176:Protein Biosynthesis / Q000187:drug effects; D011808:Quinone Reductases / Q000096:biosynthesis*; D051381:Rats; D013347:Subcellular Fractions / Q000201:enzymology
D003574:Cytochrome c Group; D046911:Macromolecular Substances; D009097:Multienzyme Complexes; D002701:Chloramphenicol; D003513:Cycloheximide; D009247:NADH, NADPH Oxidoreductases; D011808:Quinone Reductases; D003576:Electron Transport Complex IV; D014450:Electron Transport Complex III