Study of tissue-specific dihydrodiol dehydrogenase

组织特异性二氢二醇脱氢酶的研究

• 批准号: 11672175
• 负责人: HARA Akira
• 金额: $ 2.3万
• 依托单位: Gifu Pharmaceutical University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11672175/
• 关键词: dihydrodiol dehydrogenase; D-xylose dehydrogenase; new protein family; structure-function relationship; site-directed mutagenesis; anti-inflammatory drug; gene structure; 新規蛋白質ファミリー

Dihydrodiol dehydrogenase (DD, EC 1.3.1.20), which catalyzes the NADP-dependent oxidation of trans-dihydrodiols of aromatic hydrocarbons, has been shown to be involved in cytotoxicity of the aromatic hydrocarbons. The enzyme exits in multiple forms in mammalian tissues. Of the multiple forms dimeric DD is tissue-specifically expressed, but its structure and physiological function have not been elucidated. This study was performed to elucidate the structure of dimeric DD, its relationship to the function and possible clinical significance. The results obtained are summarized.1. Dimeric DD in monkey kidney, pig and dog liver was demonstrated to be identical with NADP-dependent D-xylose dehydrogenase (EC 1.1.1.179).2. The protein sequencing and/or cDNA cloning primary structures of dimeric DDs in rabbit lens, human intestine and the above animal tissues revealed that the enzymes constitute a new protein family with prokaryotic proteins including glucose-fructose oxidoreductase.3. The site … More -directed mutagenesis on the monkey dimeric DD suggested that Tyr-180 is a catalytic residue, Lys-97 and His-79 function both coenzyme binding and chemical steps of the reaction, and Tyr-71 and His-76 are involved in coenzyme binding. In addition, Trp-125, Asp-176 and Trp-254 were suggested to be responsible for substrate binding, and of the three residues Asp-176 is thought to be important for the adaptation of the alcohol substrate in to the binding site. The crystallographic study is now in progress.4. Dimeric DD was inhibited by several drugs such as nonsteroidal anti-inflammatory agents, of which the potent inhibitors commonly had 4-hydroxyphenylketone structure. The enzyme's mRNA, although it did not detected in normal human kidney, was detected in one of five renal cancer specimen. The variant gene with respect to exon 4 region was not detected, and such a study for other exon regions will be continued to find clinical significance.5. During the course of cloning of the genomic gene for dimeric, I noticed that the gene is included in a working draft sequence of chromosome 19. The 5'-noncoding region of this gene contains various putative binding sites for transcription factors.6. The structural and functional aspects obtained in this study was presented in 10^<th> International Symposium on Enzymology and Molecular Biology of Carbonyl Metabolism (New Mixico, U.S.A., July, 2000). Less

二氢二醇脱氢酶（DD，EC 1.3.1.20）催化了芳族碳氢化合物的反二氢二醇的NADP依赖性氧化，已被证明与芳族碳氢化合物的细胞毒性有关。这些酶在哺乳动物组织中以多种形式退出。多种形式的二聚体DD是特定表达的，但尚未阐明其结构和身体功能。进行这项研究是为了阐明二聚体DD的结构，其与功能的关系和可能的临床意义。获得的结果总结了1。猴肾，猪和狗肝脏中的二聚体DD与NADP依赖性的D- Xylose脱氢酶相同（EC 1.1.1.179）.2。兔晶状体，人肠和上述动物组织中二聚体DDS的蛋白测序和/或cDNA克隆的原代结构表明，酶构成了一种新的蛋白质家族，其中具有原核生物蛋白，包括葡萄糖 - 果糖果糖氧化还原酶。3。该地点……在猴子二聚体DD上进行的较为指导的诱变表明，Tyr-180是催化居住区，Lys-97和His-79功能辅酶结合和反应的化学步骤，而Tyr-71和His-76涉及辅酶结合。此外，建议认为TRP-​​125，ASP-176和TRP-254负责底物结合，而在三个保留的ASP-176中，ASP-176被认为对于在结合位点适应酒精基质的适应很重要。晶体学研究正在进行中4。二聚体DD被多种药物（例如非甾体抗炎剂）抑制，其中有效抑制剂通常具有4-羟基苯基酮结构。该酶的mRNA虽然在正常肾脏中未检测到，但在五个肾癌标本之一中被发现。未检测到相对于外显子4区域的变异基因，对其他外显子区域的这项研究将继续发现临床意义。5。在二聚体基因组基因的克隆过程中，我注意到该基因被包含在19号染色体的工作草案序列中。该基因的5'-非编码区域包含转录因子的各种推定结合位点。6。这项研究中获得的结构和功能方面在羰基代谢的酶学和分子生物学的10^<th>中提出（美国新米斯科，2000年7月）。较少的

项目成果

E.Arimitsu et al.: "Cloning and sequencing of the cDNA species for mammalian dihydrodiol dehydrogenases"Biochem.J.. 342・3. 721-728 (1999)

E.Arimitsu等：“哺乳动物二氢二醇脱氢酶的cDNA种类的克隆和测序”Biochem.J. 342·3(1999)。

S.Aoki et al.: "Identity of dimeric dihydrodiol dehydrogenase as NADP^+-dependent D-xylose dehydrogenase in pig liver"Chemico-Biol.Interact.. (in the press). (2001)

S.Aoki等人：“猪肝中二聚二氢二醇脱氢酶作为NADP 2 -依赖性D-木糖脱氢酶的身份”Chemico-Biol.Interact..（出版中）。

S.Aoki et al.: "Identity of dimeric dihydrodiol dehydrogenase as NADP^+-dependent D-xylose dehydrogenase in pig liver"Chemico-Biol.Interact.. (印刷中). (2001)

S. Aoki 等人：“猪肝中二聚二氢二醇脱氢酶作为 NADP + 依赖性 D-木糖脱氢酶的身份”Chemico-Biol.Interact..（印刷中）。

Y.Asada et al.: "Roles of His-79 and Tyr-180 of D-xylose/dihydrodiol Dehydrogenase in catalytic function"Biochem.Biophys.Res.Commun.. 278-2. 333-337 (2000)

Y.Asada 等：“D-木糖/二氢二醇脱氢酶的 His-79 和 Tyr-180 在催化功能中的作用”Biochem.Biophys.Res.Commun. 278-2。