[Affinity chromatography of menadione reductase].
The preparation of affinity adsorbents for menadione reductase (EC 1.6.99.2), using CNBr-activated Sepharose 4B and activated CH-Sepharose 4B, is described. The adsorbents used are the sepharoses modified by 4-anilino-5-methoxybenzoquinone-1,2. Bound menadione reductase is specifically eluted with NAD(P)H, while NAD(P) has no effect on the enzyme. The enzyme yield of 83--95% and a 102-fold purification are achieved. Disc-electrophoresis reveals one protein fraction identified with menadione reductase and a separate inactive fraction. It is assumed that the specific binding of menadione reductase to the adsorbent is due to the formation of a complex between the oxidized enzyme and the quinone. This complex breaks down in the presence of NAD(P)H.
D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods; D004589:Electrophoresis, Disc; D008099:Liver / Q000201:enzymology; D009247:NADH, NADPH Oxidoreductases / Q000302:isolation & purification*; D011808:Quinone Reductases / Q000302:isolation & purification*; D051381:Rats
D009247:NADH, NADPH Oxidoreductases; D011808:Quinone Reductases
Affinnaia khromatografiia menadionreduktazy.