Open AccessAffinity chromatography purification of cytochrome c binding enzymes.
Author(s) -
Angelo Azzi,
Kurt Bill,
Clemens Broger
Publication year - 1982
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.79.8.2447
Subject(s) - cytochrome c peroxidase , cytochrome c oxidase , affinity chromatography , biochemistry , cytochrome p450 reductase , chemistry , cytochrome c , reductase , cytochrome b , cytochrome , cytochrome c1 , peroxidase , enzyme , coenzyme q – cytochrome c reductase , mitochondrion , mitochondrial dna , gene
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.