Chicken heart ubiquinol: cytochrome c reductase--isolation by affinity chromatography and resolution of enzyme into subunits.
Chicken heart mitochondrial ubiquinol: cytochrome c reductase has been isolated as a monodisperse form by solubilization with Triton X-100 in low ionic strength and subsequent affinity chromatography. The enzyme complex contains less than 0.15 mumol ubiquinone and 12 mumol phospholipid per mumol enzyme. The spectrum of cytochrome b in the reduced exhibited two maximum alpha-absorption bands at 558 nm and 562 nm; a shoulder at 553 nm for c1. The purified enzyme complex consists of at least 9 subunits with apparent molecular weight of 47 K, 44 K, 36 K, 31 K, 25 K, 17 K, 15 K, 12 K and 9 K. The enzyme complex has been dissociated into subdomains of subunits in the presence of increasing concentration of salt and Triton X-100. Three subunits carrying the redox centers (cytochrome b, cytochrome c1 and Fe-S protein) have been further isolated.
D000818:Animals; D002645:Chickens / Q000378:metabolism*; D002846:Chromatography, Affinity; D003573:Cytochrome b Group / Q000302:isolation & purification; D014450:Electron Transport Complex III; D008929:Mitochondria, Heart / Q000201:enzymology*; D009097:Multienzyme Complexes / Q000302:isolation & purification* / Q000378:metabolism; D011808:Quinone Reductases / Q000302:isolation & purification* / Q000378:metabolism
D003573:Cytochrome b Group; D009097:Multienzyme Complexes; D011808:Quinone Reductases; D014450:Electron Transport Complex III