Hyndman D J, Takenoshita R, Vera N L, Pang S C, Flynn T G
Department of Biochemistry, Queen's University, Kingston, Ontario, K7L 3N6 Canada.
J Biol Chem. 1997 May 16;272(20):13286-91. doi: 10.1074/jbc.272.20.13286.
Treatment of Chinese hamster ovary (CHO) cells by the aldehyde containing calpain inhibitor I resulted in the induction of a 35-kDa protein that was partially sequenced and shown to be a member of the aldo-keto reductase superfamily (Inoue, S., Sharma, R. C., Schimke, R. T., and Simoni, R. D. (1993) J. Biol. Chem. 268, 5894-5898). Using rapid amplification of cDNA ends polymerase chain reaction, we have sequenced the cDNA for this protein (CHO reductase). This enzyme is a new member of the aldo-keto reductase superfamily and shows greatest amino acid sequence identity to mouse fibroblast growth factor-regulated protein and mouse vas deferens protein (92 and 80% sequence identity, respectively). The enzyme exhibits about 70% sequence identity with the aldose reductases (ALR2; EC 1.1.1.21) and about 47% with the aldehyde reductases (ALR1; EC 1.1.1.2). Northern analysis showed that it is induced in preference to either ALR1 or ALR2 and RNase protection assays showed gene expression in bladder, testis, jejunum, and ovary in descending order of expression. The cDNA for this inducible reductase was cloned into the pET16b vector and expressed in BL21(DE3) cells. Expressed CHO reductase showed kinetic properties distinct from either ALR1 or ALR2 including the ability to metabolize ketones. This protein joins a growing number of inducible aldo-keto reductases that may play a role in cellular regulation and protection.
用含醛基的钙蛋白酶抑制剂I处理中国仓鼠卵巢（CHO）细胞，可诱导产生一种35 kDa的蛋白质，对其进行了部分测序，结果表明它是醛酮还原酶超家族的成员之一（井上，S.，夏尔马，R.C.，施姆克，R.T.，以及西蒙尼，R.D.（1993年）《生物化学杂志》268卷，5894 - 5898页）。利用cDNA末端快速扩增聚合酶链反应，我们对该蛋白质（CHO还原酶）的cDNA进行了测序。这种酶是醛酮还原酶超家族的一个新成员，与小鼠成纤维细胞生长因子调节蛋白和小鼠输精管蛋白的氨基酸序列同一性最高（分别为92%和80%的序列同一性）。该酶与醛糖还原酶（ALR2；EC 1.1.1.21）的序列同一性约为70%，与醛还原酶（ALR1；EC 1.1.1.2）的序列同一性约为47%。Northern分析表明，它比ALR1或ALR2更易被诱导，核糖核酸酶保护分析表明该基因在膀胱、睾丸、空肠和卵巢中的表达呈递减顺序。将这种可诱导还原酶的cDNA克隆到pET16b载体中，并在BL21(DE3)细胞中表达。表达的CHO还原酶表现出与ALR1或ALR2不同的动力学特性，包括代谢酮的能力。这种蛋白质加入了越来越多可能在细胞调节和保护中发挥作用的可诱导醛酮还原酶的行列。
