Morjana N A, Flynn T G
Department of Biochemistry, Queen's University, Kingston, Ontario, Canada.
J Biol Chem. 1989 Feb 15;264(5):2906-11.
Aldose reductase (ALR2) has been purified to homogeneity from human psoas muscle. From sodium dodecyl sulfate-polyacrylamide electrophoresis the enzyme is monomeric and has a molecular weight of 37,000. ALR2 catalyzes the primarily NADPH-dependent reduction of a wide variety of aldehydes, although the enzyme can also utilize NADH. The best substrates for ALR2 are aromatic aldehydes (e.g. pyridine-3-aldehyde; Km = 9 microM; kcat/Km = 150,000 s-1 M-1), while among aldoses DL-glyceraldehyde is the preferred substrate (Km = 72 microM; kcat/Km = 17,250). Low (100 microM) concentrations of CaCl2 and CaSO4 cause a marked inhibition (90%) of ALR2 as do higher concentrations (0.2 M) of MgCl2. (NH4)2SO4 caused a 2-fold activation of ALR2. The enzyme is also inhibited by quercetin and the commercially developed aldose reductase inhibitors alrestatin and sorbinil. ALR2 is inhibited only very slightly by sodium valproate and barbiturates. ALR2 cross-reacts immunologically with human brain and human placental aldose reductase and with ALR2 from monkey tissue. There is no precipitin cross-reaction of ALR2 with aldose reductases from other species nor with human aldehyde reductase 1 (ALR1) or with ALR1 from other species. The data show that human muscle is a new and relatively rich source of a monomeric NADPH/NADH reductase which is clearly identifiable as aldose reductase.
已从人腰大肌中纯化出均一的醛糖还原酶(ALR2)。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳可知,该酶为单体,分子量为37,000。ALR2主要催化依赖NADPH的多种醛的还原反应,不过该酶也能利用NADH。ALR2的最佳底物是芳香醛(如吡啶-3-醛;Km = 9微摩尔;kcat/Km = 150,000 s-1 M-1),而在醛糖中,DL-甘油醛是首选底物(Km = 72微摩尔;kcat/Km = 17,250)。低浓度(100微摩尔)的CaCl2和CaSO4会对ALR2产生显著抑制(90%),高浓度(0.2 M)的MgCl2也是如此。硫酸铵可使ALR2活性提高2倍。该酶也会受到槲皮素以及商业开发的醛糖还原酶抑制剂阿雷司他汀和索比尼尔的抑制。丙戊酸钠和巴比妥类药物对ALR2的抑制作用非常轻微。ALR2与人脑、人胎盘醛糖还原酶以及猴组织中的ALR2发生免疫交叉反应。ALR2与其他物种的醛糖还原酶、人醛糖还原酶1(ALR1)或其他物种的ALR1之间不存在沉淀素交叉反应。数据表明,人肌肉是一种新的、相对丰富的单体NADPH/NADH还原酶来源,该还原酶可明确鉴定为醛糖还原酶。