Characterization of hamster NAD+-dependent 3(17)β-hydroxysteroid dehydrogenase belonging to the aldo-keto reductase 1C subfamily.
Characterization of hamster NAD+-dependent 3(17)β-hydroxysteroid dehydrogenase belonging to the aldo-keto reductase 1C subfamily.
复制标题
属于醛酮还原酶 1C 亚家族的仓鼠 NAD+ 依赖性 3(17)β-羟基类固醇脱氢酶的表征。
DOI：
10.1093/jb/mvv057
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发表时间：
2015
影响因子：
2.7
通讯作者：
T. Matsunaga
中科院分区：
文献类型：
--
作者：
S. Endo;Misato Noda;A. Ikari;K. Tatematsu;O. El;A. Harã;Y. Kitade;T. Matsunaga
从金黄地鼠肝中克隆了吗啡6-脱氢酶（AKR 1C 34）及其同源醛酮还原酶（AKR 1C 35）的cDNA，并比较了它们的酶学性质和组织分布。AKR 1C 34和AKR 1C 35类似地使用NAD（+）氧化各种异生物质脂环醇，但它们对羟基类固醇的底物特异性和抑制剂敏感性不同。AKR 1C 34显示3α/17β/20α-羟基类固醇脱氢酶活性，AKR 1C 35有效氧化各种3β-和17β-羟基类固醇，包括具有生物活性的3β-羟基-5 α/β-二氢-C19/C21-类固醇、脱氢表雄酮和17β-雌二醇。AKR 1C 35与AKR 1C 34的不同之处还在于其对类黄酮的高敏感性，类黄酮与17β-雌二醇竞争性抑制（Ki 0.11-0.69 μM）。AKR 1C 35的mRNA在雄性仓鼠中表达为肝脏特异性的，在雌性仓鼠中普遍存在，而AKR 1C 34的mRNA的表达显示出相反的性别二型性。由于AKR 1C 35是AKR超家族中的第一个317Β-羟基类固醇脱氢酶，我们还通过将AKR 1C 35中的Val 54和Cys 310分别替换为AKR 1C 34中的相应残基Ala和Phe，研究了3β-羟基类固醇脱氢酶活性的分子决定因素。Val 54 Ala的突变，而不是Cys 310 Phe，显着损害这种活性，表明Val 54在识别类固醇底物中起着关键作用。
The cDNAs for morphine 6-dehydrogenase (AKR1C34) and its homologous aldo-keto reductase (AKR1C35) were cloned from golden hamster liver, and their enzymatic properties and tissue distribution were compared. AKR1C34 and AKR1C35 similarly oxidized various xenobiotic alicyclic alcohols using NAD(+), but differed in their substrate specificity for hydroxysteroids and inhibitor sensitivity. While AKR1C34 showed 3α/17β/20α-hydroxysteroid dehydrogenase activities, AKR1C35 efficiently oxidized various 3β- and 17β-hydroxysteroids, including biologically active 3β-hydroxy-5α/β-dihydro-C19/C21-steroids, dehydroepiandrosterone and 17β-estradiol. AKR1C35 also differed from AKR1C34 in its high sensitivity to flavonoids, which inhibited competitively with respect to 17β-estradiol (Ki 0.11-0.69 μM). The mRNA for AKR1C35 was expressed liver-specific in male hamsters and ubiquitously in female hamsters, whereas the expression of the mRNA for AKR1C34 displayed opposite sexual dimorphism. Because AKR1C35 is the first 317Β-HYDROXYSTEROID DEHYDROGENASE IN THE AKR SUPERFAMILY: , we also investigated the molecular determinants for the 3β-hydroxysteroid dehydrogenase activity by replacement of Val54 and Cys310 in AKR1C35 with the corresponding residues in AKR1C34, Ala and Phe, respectively. The mutation of Val54Ala, but not Cys310Phe, significantly impaired this activity, suggesting that Val54 plays a critical role in recognition of the steroidal substrate.
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影响因子：
--
作者：
Reddy, Doodipala Samba
通讯作者：
Reddy, Doodipala Samba
影响因子：
20.3
作者：
T. Penning
通讯作者：
T. Penning
影响因子：
2.9
作者：
Jin, Y;Stayrook, SE;Lewis, M
通讯作者：
Lewis, M
影响因子：
4.8
作者：
Steckelbroeck, S;Jin, Y;Penning, TM
通讯作者：
Penning, TM
DOI：
--
发表时间：
2000
期刊：
The Biochemical journal
影响因子：
--
作者：
T. Penning;M. Burczynski;J. Jez;C. Hung;H. K. Lin;H. Ma;M. Moore;N. Palackal;K. Ratnam
通讯作者：
T. Penning;M. Burczynski;J. Jez;C. Hung;H. K. Lin;H. Ma;M. Moore;N. Palackal;K. Ratnam