Structure and Function of NAD/NADP Requirement Microsomal Alcohol Dehydrogensase

NAD/NADP 需求微粒体乙醇脱氢酶的结构和功能

• 批准号: 10672068
• 负责人: MATSUNAGA Tamihide
• 金额: $ 1.92万
• 依托单位: Hokuriku University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10672068/
• 关键词: 3-OH-β-ionone; 3-oxo-β-ionone; alcohol; ketone; microsomes; dehydrogenase; oxidation; NAD(P); 脱水素酸化; NAD; NADP; サル

1. 3-OH-β-ionone was oxidized to 3-oxo-β-ionone by hepatic microsomes of mouse and monkey. In both animals, NAD was most effective as cofactor, followed by NADP, NADPH and NADH.2. The NADPH-dependent 3-oxo-β-ionone forming activity (MALCO activity) in mouse was almost completely inhibited by antibody against Cyp3all.3-OH-β-ionone MALCO activity in monkey was inhibited by antisera against CYP2A and Cyp3all.4. The NAD- and NADP-dependent formation of 3-oxa-β-ionone in mouse was inhibited by disulfiram.5. In the case of monkey, the NADP-dependent activity was inhibited by babital, quercetin and quercitrin as well as disulfiram and HgClィイD22ィエD2, while the NAD-dependent activity was inhibited by disulfiram and HgClィイD22ィエD2.6. In the hepatic microsomes of monkey, the Km/Vmax value for NAD-dependent 3-oxo-β-ionone forming activity was about 10-fold higher than that for the NADPH-dependent activity.7. Microsomal 3-OH-β-ionone dehydrogenate (MALDH) was purified from male Japanese monkey livers. Specific activity of the enzyme was about 5 times higher than that of microsomes, and the molecular mass is about 23.5 kDa.These results indicated that not only P450(MALCO) but also microsomal alcohol dehydrogenase(s) (MALDH) required NAD and/or NADP as cofactor catalyzed the formation of 3-oxo-β-ionone from 3-OH-β-ionone in hepatic microsomes of mouse and monkey.

1. 3-OH-β-紫罗兰酮被小鼠和猴的肝微粒体氧化为3-氧代-β-紫罗兰酮。在这两种动物中，NAD 作为辅助因子最有效，其次是 NADP、NADPH 和 NADH。2。小鼠中 NADPH 依赖性 3-oxo-β-紫罗兰酮形成活性（MALCO 活性）几乎完全被 Cyp3all 抗体抑制。CYP2A 和 Cyp3all 抗血清抑制猴中 3-OH-β-紫罗兰酮 MALCO 活性。4。双硫仑可抑制小鼠体内 NAD 和 NADP 依赖性 3-氧杂-β-紫罗兰酮的形成。5．在猴的情况下，NADP依赖性活性被巴比妥、槲皮素和槲皮素以及双硫仑和HgClィイD22ィエD2抑制，而NAD依赖性活性被双硫仑和HgClィイD22ィエD2.6抑制。在猴肝微粒体中，NAD依赖性3-氧代-β-紫罗兰酮形成活性的Km/Vmax值比NADPH依赖性活性高约10倍。7．从雄性日本猴肝脏中纯化微粒体 3-OH-β-紫罗兰酮脱氢物 (MALDH)。该酶的比活性比微粒体高约5倍，分子量约为23.5 kDa。这些结果表明，不仅P450（MALCO）而且微粒体乙醇脱氢酶（MALDH）都需要NAD和/或NADP作为辅助因子，催化肝内3-OH-β-紫罗兰酮形成3-oxo-β-紫罗兰酮。 小鼠和猴子的微粒体。

项目成果

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