Catalytic activity and arrangement of subunit polypeptides in rat liver cytochrome c oxidase as studied by proteolysis.
Cytochrome c oxidase from rat liver was incubated with various proteinases of different specificities and the enzymic activity was measured after various incubation times. A loss of catalytic activity was found after digestion with proteinase K, aminopeptidase M and a mitochondrial proteinase from rat liver. In each case the decrease in enzymic activity was compared with the changes in intensities of the polypeptide pattern obtained after sodium dodecyl sulfate polyacrylamide gel electrophoresis. The susceptibilities of the subunit polypeptides of the soluble cytochrome c oxidase to proteinases were very different. Whereas subunit I was most susceptible, subunits V--VII were rather resistant to degradation. From the relative inaccessibility of subunits V--VII to proteinases it is likely that these polypeptides are buried in the interior of the enzyme complex.
D000626:Aminopeptidases / Q000378:metabolism; D000818:Animals; D003840:Deoxycholic Acid; D003576:Electron Transport Complex IV; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D046911:Macromolecular Substances; D008928:Mitochondria / Q000201:enzymology*; D008930:Mitochondria, Liver / Q000201:enzymology*; D010447:Peptide Hydrolases / Q000378:metabolism; D051381:Rats; D013367:Submitochondrial Particles / Q000201:enzymology*
D046911:Macromolecular Substances; D003840:Deoxycholic Acid; D003576:Electron Transport Complex IV; D010447:Peptide Hydrolases; D000626:Aminopeptidases