ANALYSIS OF GENE MUTATION IN THAI CAUSING FISH ODOR SYNDROME

泰国鱼臭综合症的基因突变分析

• 批准号: 10044224
• 负责人: KAMATAKL Tetsuya
• 金额: $ 4.93万
• 依托单位: HOKKAIDO UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A).
• 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-10044224/
• 关键词: trimethylamine; FMO; genetic polymorphism; genetic disease; gene therapy; heterologous expression; human genome; gene analysis; ノックアウトマウス; クローニング; 代謝欠損; 発現系; 遺伝子多型

We examined the metabolism of trimethylamine (TMA) in human liver microsomes. The kinetic parameters of TMA N-oxidation showed the apparent Km and Vmax values of 45.9 μM and 2.24 nmol/min/mg protein, respectively. The N-oxidation of TMA was inhibited by the known flavin-containing monooxygenase (FMO) inhibitors such as methimazole and heat treatment, but not by SKF-525A, cytochrome P450 inhibitor. To identify the FMO form responsible for the N-oxidation of TMA, the TMA N-oxidase activities of human FMO1, FMO3 and FMO5 expressed in E, coli were measured. FMO5 showed no TMA N-oxidase activity. The Vmax/Km value for FMO3 was about 355 times higher than that for FMO1 when the kinetic parameters of TMA N-oxidation was examined. We analyzed the FMO3 gene of Thai possible patients with fish-odor syndrome. A novel mutation, a single base substitution from G to A at the position of 265 (G265A), was identified in exon 3 of two patients. The mutated FMO3 protein with an amino acid substitution from valine to isoleucine at residue 58 (V58I) exhibited the reduced TMA N-oxidase activity when it was expressed in Salmonella. We conclude that FMO3 gene with this mutation produce the FMO protein that has reduced catalytic activities.

我们研究了三甲胺（TMA）在人肝微粒体中的代谢。TMA N-氧化的动力学参数显示表观Km和Vmax值分别为45.9 μM和2.24 nmol/min/mg蛋白。甲巯咪唑和热处理等已知的含黄素单加氧酶（FMO）抑制剂可抑制TMA的N-氧化，而细胞色素P450抑制剂SKF-525 A则不抑制TMA的N-氧化。为了鉴定负责TMA的N-氧化的FMO形式，测量了在大肠杆菌中表达的人FMO 1、FMO 3和FMO 5的TMA N-氧化酶活性。FMO 5没有表现出TMA N-氧化酶活性。当考察TMA N-氧化的动力学参数时，FMO 3的Vmax/Km值比FMO 1高约355倍。我们分析了泰国可能患有鱼腥味综合征的患者的FMO 3基因。在2例患者的第3外显子中发现了一个新的突变，即第265位由G变为A（G265 A）。在第58位氨基酸残基（V58 I）由缬氨酸替换为异亮氨酸的突变FMO 3蛋白在沙门氏菌中表达时表现出降低的TMA N-氧化酶活性。我们的结论是，FMO 3基因与此突变产生的FMO蛋白的催化活性降低。

项目成果

鎌滝哲也: "薬物療法学"南山堂 (in press).

Tetsuya Kamataki：“药物治疗学”Nanzando（正在出版）。

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Ken-ichi Nunoya 等人：“CYP2A6 基因缺失负责人体内 ( )-cis-3,5-二甲基-2-(3-吡啶基)噻唑烷-4-酮盐酸盐的体内多态性代谢”药理学杂志

Taisei Mushiroda, Noritaka Ariyoshi, Kanzo Kimura, Eiji Kashiyama, Osamu Nagata, Hideo Kato, and Tetsuya Kamataki: "A high level expression of CYP2A in the lung of the Suncus (Suncus murinus) and its role in the Activation of 4-(methylntirosamino)-1-(3-py

Taisei Mushiroda、Noritaka Ariyoshi、Kanzo Kimura、Eiji Kashiyama、Osamu Nagata、Hideo Kato 和 Tetsuya Kamataki：“CYP2A 在 Suncus (Suncus murinus) 肺部的高水平表达及其在 4-(methylntirosamino) 激活中的作用

Taisei Mushiroda, et al.: "The Suncus (Suncus murlnus) shows poor metabolic phenotype for trimethylamine N-oxygenation"Toxicology and Applied Pharmacology. In press.

Taisei Mushiroda 等人：“Suncus (Suncus murlnus) 对于三甲胺 N-氧化表现出较差的代谢表型”毒理学和应用药理学。

鎌滝哲也他: "環境要因感受性を支配する遺伝的要因“環境と健康"" へるす出版, 245-29 (1998)

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