Affinity chromatography purification of cytochrome c binding enzymes.
An efficient affinity chromatography procedure for the isolation of mitochondrial cytochrome c oxidase and reductase is described. Saccharomyces cerevisiae cytochrome c was used as a ligand, bound to a thiol-Sepharose 4B gel through cysteine-107. In this way, the site of interaction of cytochrome c with cytochrome oxidase and reductase remained unmodified and available for binding to a number of partner enzymes. The procedure is adequate for the purification of all those proteins having in common the property of binding with high affinity to cytochrome c--e.g., cytochrome c oxidase, reductase, and peroxidase, sulfite oxidase, and reaction centers of photosynthetic bacteria.
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods; D003579:Cytochrome Reductases / Q000302:isolation & purification*; D003574:Cytochrome c Group; D003578:Cytochrome-c Peroxidase / Q000302:isolation & purification*; D003580:Cytochromes / Q000302:isolation & purification*; D003576:Electron Transport Complex IV / Q000302:isolation & purification*; D006736:Horses; D008929:Mitochondria, Heart / Q000201:enzymology; D009206:Myocardium; D009245:NADH Dehydrogenase / Q000302:isolation & purification*; D010088:Oxidoreductases / Q000302:isolation & purification*; D050862:Oxidoreductases Acting on Sulfur Group Donors / Q000302:isolation & purification*; D010544:Peroxidases / Q000302:isolation & purification*; D011487:Protein Conformation; D012441:Saccharomyces cerevisiae
D003574:Cytochrome c Group; D003580:Cytochromes; D010088:Oxidoreductases; D010544:Peroxidases; D003578:Cytochrome-c Peroxidase; D003579:Cytochrome Reductases; D009245:NADH Dehydrogenase; D050862:Oxidoreductases Acting on Sulfur Group Donors; D003576:Electron Transport Complex IV