Molecular cloning of testicular 20 alpha-hydroxysteroid dehydrogenase: identity with aldose reductase.
Molecular cloning of testicular 20 alpha-hydroxysteroid dehydrogenase: identity with aldose reductase.
复制标题
睾丸 20 α-羟基类固醇脱氢酶的分子克隆：与醛糖还原酶的同一性。
DOI：
10.1021/bi00057a003
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发表时间：
1993
期刊：
影响因子：
2.9
通讯作者：
Zimmer,WE
中科院分区：
文献类型：
--
作者：
Warren,JC;Murdock,GL;Ma,Y;Goodman,SR;Zimmer,WE
1992年11月23日收到的摘要：使用抗20 α-羟基类固醇脱氢酶（20 α-HSD）的多克隆抗体和DNA探针杂交，从牛睾丸Xgt 11文库中分离出编码20 α-HSD的互补DNA（cDNA）克隆。使用三个独立分离的克隆的核苷酸测序来建立编码该酶的复合cDNA序列。它含有921个核苷酸的编码序列，一个终止密码子和一个264个核苷酸的3/-非编码片段，该片段允许推导该酶的氨基酸序列。对GenBank DNA序列数据库进行的20 α-HSD cDNA的计算机同源性搜索显示其与牛透镜醛糖还原酶（alditohNADPH氧化还原酶; EC 1.1. 1.21），文献检索显示推导的氨基酸序列与报道的牛酶相同。从纯化的睾丸20 α-HSD的N-末端和从用内肽酶Lys-C处理的随机肽获得的序列都与20 α-HSD的推导的氨基酸序列和/或公开的醛糖还原酶序列的区域相同。此外，该酶纯化后，通过以下活动与17-羟孕酮作为底物的同质性被证明是减少葡萄糖，甘油醛，苯甲醛（所有经典的醛糖还原酶底物）。最后，17-羟孕酮抑制苯甲醛和甘油醛的还原。Shikita及其同事报道了在大鼠和猪睾丸中存在20 α-羟基类固醇脱氢酶（20 α-HSD）1（Shikita et al.，1967; Sato等人，1972年）。该酶为单体酶（MT 35000），对雌酮无170-羟基类固醇脱氢酶（170-HSD）活性。国际生物化学联合会命名委员会推荐将这种酶命名为20 α-羟类固醇脱氢酶，并将其编号为EC 1.1。1.149（提名委员会，1979年）。在从事设计用于确定类固醇结合位点的拓扑结构和哺乳动物类固醇互变酶的催化机制的研究时，我们使人胎盘170-羟基类固醇脱氢酶结晶（Chin等人，1976年），并确定了类固醇结构域和某些
Revised Manuscript Received November 23, 1992 abstract: Complementary DNA (cDNA) clones encoding bovine testicular 20a-hydroxysteroid dehydrogenase (20a-HSD) have been isolated from a bovine testicular Xgtl 1 library using polyclonal antibodies against 20a-HSD and DNA probe hybridization. Nucleotide sequencing of three independently isolated clones was used to establish a composite cDNA sequence that encodes the enzyme. It contains a coding sequence of 921 nucleotides, a stop codon, and a 264-nucleotide 3/-noncoding segment which allowed deduction of the amino acid sequence of the enzyme. A computer homology search of the 20a-HSD cDNA performed against the GenBank DNA sequence database revealed it to be identical with bovine lens aldose reductase (alditohNADPH oxidoreductase; EC 1.1. 1.21), and a literature search revealsthe deduced amino acid sequence to be identical with that reported for the bovineenzyme. Sequences obtainedfrom the N-terminus of purified testicular 20a-HSD and from random peptides obtained by treatment with endopeptidase Lys-C are all identical with regions of the deduced amino acid sequence of 20a-HSD and/or the published sequence of aldose reductase. Further, theenzyme purified to homogeneity by following activity with 17-hydroxyprogesterone as a substrate was shown to reduce glucose, glyceraldehyde, and benzaldehyde (all classic aldose reductase substrates). Finally, 17-hydroxyprogesterone inhibited the reduction of benzaldehyde and glyceraldehyde. Because aldose reductase has been implicatedin the etiology of diabetic complications, acceptance of steroid substrates may offer new implications for therapy.Shikita and co-workers reported the presence of a 20ahydroxysteroid dehydrogenase (20a-HSD) 1 in rat and porcine testis (Shikita et al., 1967; Sato et al., 1972). The enzyme was shown to bea monomer(MT 35 000) and tolack 170-hydroxysteroid dehydrogenase (170-HSD) activity toward estrone. TheNomenclature Committee of the International Union of Biochemistry recommended the name 20a-hydroxysteroid dehydrogenase for this enzyme andassigned it the number EC 1.1. 1.149 (Nomenclature Committee, 1979). While engaged in studies designed to define the topography of the steroid binding site and the catalytic mechanisms of mammalian steroid-interconverting enzymes, we had crys-tallized human placental 170-hydroxysteroid dehydrogenase (Chin et al., 1976) and identified the steroid domain and certain