大鼠肝脏醛还原酶的分离与鉴定

Isolation and characterization of rat liver aldehyde reductase.

作者信息

Turner A J, Hryszko J

出版信息

Biochim Biophys Acta. 1980 Jun 13;613(2):256-65. doi: 10.1016/0005-2744(80)90081-9.

Abstract

A systematic investigation of potential ligands for the affinity purification of aldehyde reductase (alcohol:NADP+ oxidoreductase, EC 1.1.1.2.) has been carried out. The most suitable nucleotide ligands tested were NADP+ and 2',5'-ADP. Adsorbed enzyme could be eluted with NADPH but not NADH. The chlorotriazinyl dyes Cibacron Blue F3GA and Procion Red HE3B also proved effective as 'affinity' ligands when immobilized to Sepharose 4B. The free dyes and also Blue Dextran (Cibacron Blue F3GA coupled to dextran) were all potent inhibitors of aldehyde reductase. The inhibition by Blue Dextran was shown to be competitive with respect to NADPH (Ki = 1.8 x 10(-7) M). The enzyme was sensitive to inhibition by glutaric acid derivatives, flavonoids and a range of anti-convulsants.

摘要

已对用于醛还原酶（醇：NADP +氧化还原酶，EC 1.1.1.2.）亲和纯化的潜在配体进行了系统研究。测试的最合适的核苷酸配体是NADP +和2'，5'-ADP。吸附的酶可用NADPH洗脱，但不能用NADH洗脱。当固定在琼脂糖4B上时，氯三嗪基染料汽巴蓝F3GA和普施安红HE3B也被证明是有效的“亲和”配体。游离染料以及蓝色葡聚糖（与葡聚糖偶联的汽巴蓝F3GA）都是醛还原酶的有效抑制剂。已表明蓝色葡聚糖的抑制作用相对于NADPH具有竞争性（Ki = 1.8×10（-7）M）。该酶对戊二酸衍生物、类黄酮和一系列抗惊厥药的抑制敏感。