[Affinity chromatography of menadione reductase on Sepharose with aminoaromatic and aminoaliphatic quinone ligands]

New modifications of Sepharoses with the ligands--4-hexamethylenimino-5-methoxy-1.2-benzoquinone (I), 4-(p-imminomethylaniline)-5-methoxy-1.2-benzoquinone (II) and 4-(p-iminomethylaniline)-5-methoxy-1.2-dihydroxybenzene (III) are described and their absorption properties with respect to menadione reductase (EC 1.6.99.2) investigated. The modification with ligand (II) affinitely adsorbs menadione reductase; the enzyme remains adsorbed in the presence of high concentration of electrolytes and within the pH range of 6.0-7.5. The elution is specific and is achieved by NAD (P) H but not by NAD (P). After chromatography the enzyme is homogeneous as shown by disc-electrophoresis in polyacrylamide gel. Under similar conditions Sepharoses with ligands (I) and (III) but poorly bind the enzyme. It is assumed that an essential role in the adsorption mechanism belongs to dispersion forces and to formation of a molecular charge-transfer complex between the aminoaromatic quinone ligand and menadione reductase active center.