[The C27-steroid hydroxylating system from bovine liver mitochondria. Isolation of ferredoxin (hepatoredoxin) by affinity chromatography on cytochrome-c-sepharose].
A technique for the isolation of highly purified hepatoredoxin involving the DE-32 cellulose chromatography of post-mitochondrial supernatant, ammonium-sulfate fractionation, Sephadex G-75 gel chromatography, 1-amino-2-hydroxypropyl-Sepharose ion-exchange chromatography and cytochrome-c-Sepharose affinity chromatography is described. The protein was purified 160-fold with a yield of 19%. The synthesis of cytochrome-c-Sepharose was carried out in a way preventing modification of the lysine-containing binding domain of the cytochrome c molecule. To achieve this, free carboxyl groups were modified with histamine to introduce imidazole residues in cytochrome c and the modified protein was immobilized on bromoacetyl-Sepharose.
D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity; D002847:Chromatography, Agarose; D002852:Chromatography, Ion Exchange; D003574:Cytochrome c Group / Q000302:isolation & purification* / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004800:Enzymes, Immobilized / Q000302:isolation & purification / Q000378:metabolism; D005288:Ferredoxins / Q000302:isolation & purification* / Q000378:metabolism; D008930:Mitochondria, Liver / Q000201:enzymology / Q000378:metabolism*; D009251:NADPH-Ferrihemoprotein Reductase / Q000302:isolation & purification / Q000378:metabolism; D013250:Steroid Hydroxylases / Q000378:metabolism*
D003574:Cytochrome c Group; D004800:Enzymes, Immobilized; D005288:Ferredoxins; D013250:Steroid Hydroxylases; D009251:NADPH-Ferrihemoprotein Reductase
C27-steroidgidroksiliruiushchaia sistema iz mitokhondriĭ pecheni byka. Vydelenie ferredoksina (gepatoredoksina) affinnoĭ khromatografieĭ na tsitokhrom-c-sefaroze.