Isolation and partial characterization of the cytochrome c oxidase of Micrococcus luteus (lysodeikticus).
The cell membrane of Micrococcus luteus (lysodeikticus) contains a respiratory chain composed of hemes a, b, and c, which contain 171, 457, and 407 pmol/mg protein, respectively. Cytochrome c oxidase, the heme a containing component, has been purified after solubilization in Triton X-100, by gel filtration on Sepharose 4B-CL ammonium sulfate precipitation and ion-exchange and affinity chromatographies on a yeast cytochrome c-Sepharose 4B column. The purified complex, which contains three polypeptides of apparent Mr 47,000, 31,000, and 19,000, has CN-sensitive ferrocytochrome c oxidase activity (Ki = 0.35 microM) and a characteristic absorption spectrum with maxima in the oxidized form at 595 and 426 nm and in the reduced form at 601 and 444 nm. The purified enzyme contains 17.4 nmol/mg protein and its copper content is 23.2 nmol/mg protein. The enzyme was purified about 100-fold with respect to its content in crude membranes. The total heme a yield, also with respect to crude membranes content, was 6.8%.
Artzatbanov
V
V
Institut für Biochemie und Molekularbiologie, Universität Bern, Switzerland.
Müller
M
M
Azzi
A
A
D002852:Chromatography, Ion Exchange; D003580:Cytochromes / Q000302:isolation & purification / Q000378:metabolism; D003576:Electron Transport Complex IV / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D008837:Micrococcus / Q000201:enzymology*
D003580:Cytochromes; D003576:Electron Transport Complex IV