title: Tissue-specific and species-specific distribution of -SH groups in cytochrome c oxidase subunits.
abstract: Cytochrome c oxidase was isolated from pig, bovine, rat and human tissues including liver, heart, diaphragm and kidney. The native and the sodium-dodecyl-sulfate (SDS)-dissociated enzymes were labelled under optimal conditions with N-ethyl-[2,3-14C]maleimide before and after reduction with dithiothreitol, separated into 13 subunits by SDS gel electrophoresis and the radioactive bands were visualized by fluorography. In some cases the radioactive bands were cut out and counted. All isozymes were labelled in subunits I, III, Va and VIIb, and in subunit II after reduction. Labelling of subunit Vb was equivocal, and in no case were subunits IV and VIc labelled. All other subunits were labelled tissue-specifically and/or species-specifically. No differences were found between labelling of the native and SDS-dissociated enzyme. By relating the molar amount of bound N-ethylmaleimide to the known amount of cysteines in subunits of bovine heart cytochrome c oxidase, the percentage of -SH group reactivity was calculated. Only the cysteine of subunit Va was found to be 100% reactive. The distinct and different reactivity of subunit VIIb as compared to subunits VIIa and VIIc clearly establishes this polypeptide as an independent subunit of mammalian cytochrome c oxidase.
mesh_terms: D000818:Animals; D002417:Cattle; D003964:Diaphragm / Q000201:enzymology; D004229:Dithiothreitol; D003576:Electron Transport Complex IV / Q000032:analysis*; D005033:Ethylmaleimide; D006801:Humans; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D009206:Myocardium / Q000201:enzymology; D009928:Organ Specificity; D051381:Rats; D013045:Species Specificity; D013438:Sulfhydryl Compounds / Q000032:analysis*; D013552:Swine
chemical_list: D013438:Sulfhydryl Compounds; D003576:Electron Transport Complex IV; D005033:Ethylmaleimide; D004229:Dithiothreitol