Horse-liver glutathione reductase: Purification and characterization

摘要: Abstract 1. Purification of horse-liver glutathione reductase was obtained by affinity chromatography on N 6 -(6-aminohexyl)-adenosine-1′5′-bisphosphate Sepharose ( -2′5′-ADP-Sepharose) and Reactive Red-120-Agarose, DEAE-Sephadex Sephacryl S-300. 2. The final preparation had 248 U/mg specific activity after 11,174-fold purification with 47% recovery, homogeneous SDS-electrophoresis. It showed charge heterogeneity in non-denaturing electrophoresis chromatofocusing, several peaks pI between 5.7 6.7. 3. enzyme homodimeric (107,000 native MW), S 20 w = 6.31 S, 41.22 A hydrodynamic radius. absorption at 270, 370 462 nm, a characteristic flavoproteins. 4. When NADPH substituted deamino-NADPH or NADH the 69 8.5% activity, respectively, while glutathione-CoA mixed disulfide 23% shown GSSG. Apparent K m values 8.8, 680, 59, 560 μ M were measured for NADPH, NADH, GSSG ferrycianide, respectively.