ALDOSE REDUCTASE: KINETICS, MECHANISM & DIABETES

醛糖还原酶：动力学、机制

• 批准号: 3230640
• 负责人: CHARLES EDWARD GRIMSHAW
• 金额: $ 10.62万
• 依托单位: SCRIPPS CLINIC AND RESEARCH FOUNDATION
• 依托单位国家: 美国
• 财政年份: 1983
• 资助国家: 美国
• 起止时间: 1983-04-01 至 1989-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3230640
• 关键词: NAD(H) phosphate; aldehyde reductase; antibody; antigens; chemical group; enzyme induction /repression; enzyme structure; enzyme substrate; gel electrophoresis; genetic mapping; glyceraldehyde 3 phosphate dehydrogenase; human tissue; interstitial nephritis; kidney metabolism; lens disorder; stereochemistry

The Action of aldose reductase (EC 1.1.1.21; "low-Km" aldehyde reductase; ALR2) is implicated in the etiology of diabetic complications of the eye, kidney, heart, and nerve. The ability of specific aldose reductase inhibitors (ARIs) to prevent the development of complications (basement membrane thickening, renal hypertrophy, defective axonal transport and nerve conduction velocity) in various tissues has confirmed the central role of ALR2 in normal and diabetic metabolism. This conclusion is supported by immunologic evidence for phylogenetic conservation of the mammalian ALR2 gene product. Homogeneous bovine kidney ALR2 (BKALR2) is virtually identical to bovine lens ALR2, but only after the BKALR2 has undergone a slow, thiol-dependent activation to a stable form. The activation process, which results in alteration of the kinetic behavior (decreased sensitivity to inhibition by Sorbinil, decreased stimulation by sulfate, appearance of non-linearity in the double-reciprocal plot for glyceraldehyde), is relevant to the activation and variable ARI sensitivity seen in human tissues. Further mechanistic studies using homogeneous BKALR2 will focus on characterization of the activation process using kinetic and immunologic techniques, identification of the physiological substrate by structure-activity studies, and determination of the mode of ARI inhibition. The extension of these studies to human kidney ALR2 and "high-Km" aldehyde reductase will utilize mon-specific rabbit anti-BKALR2 for the isolation and characterization of HKALR2. Key experiments, based on the results from the bovine system, will evaluate the mode and sensitivity to ARI inhibition, and the role of activation (as documented for BKALR2) in the human system.

醛糖还原酶的作用（EC 1.1.1.21;“low-Km”醛还原酶; ALR 2）与糖尿病眼部并发症的病因学有关， 肾脏心脏和神经 特异性醛糖还原酶 抑制剂（阿里斯），以防止并发症的发展（基础 膜增厚、肾肥大、轴突运输缺陷和 神经传导速度）在各种组织中已经证实了中枢 ALR 2在正常和糖尿病代谢中的作用。 这个结论是 支持的免疫学证据的系统发育保守的 哺乳动物ALR 2基因产物 同种牛肾ALR 2（BKALR 2）与牛肾ALR 2几乎相同。 透镜ALR 2，但只有在BKALR 2经历了缓慢的，巯基依赖性 活化成稳定的形式。 激活过程会导致 动力学行为的改变（对抑制的敏感性降低， Sorbinil，硫酸盐刺激减少， 甘油醛的双倒数图），与 在人体组织中观察到的激活和可变ARI敏感性。 进一步 使用均质BKALR 2的机制研究将集中于表征 使用动力学和免疫学技术的活化过程， 通过结构-活性鉴定生理底物 研究，并确定ARI抑制的模式。 延长 这些对人类肾脏ALR 2和“高Km”醛还原酶的研究将 利用单特异性兔抗BKALR 2进行分离， HKALR 2的特征。 关键实验，基于 牛系统，将评价ARI抑制的模式和敏感性， 以及激活（如BKALR 2所记录的）在人类系统中的作用。