MEVALONATED METABOLIZING ENZYMES & THEIR INBORN DEFECTS

甲羟酸化代谢酶

• 批准号: 6177590
• 负责人: HENRY M MIZIORKO
• 金额: $ 23.82万
• 依托单位: MEDICAL COLLEGE OF WISCONSIN
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-09-01 至 2004-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6177590
• 关键词: X ray crystallography; active sites; affinity labeling; alcohol phosphotransferase; enzyme activity; enzyme deficiency; enzyme feedback; enzyme mechanism; enzyme model; enzyme structure; fatty acid metabolism; gene mutation; isoprenoid; mevalonate; posttranslational modifications; protein sequence; recombinant proteins; site directed mutagenesis

DESCRIPTION: Mevalonic acid is a key intermediate in the biosynthesis of cellular isoprenoids and sterols. Three enzymes, mevalonate kinase, phosphomevalonate kinase, and mevalonate 5-pyrophosphate decarboxylase, are required to metabolize mevalonate to the biosynthetically active isoprenoid, isopentenyl pyrophosphate. Initial work outlined for the enzymes of mevalonate metabolism focuses on mevalonate kinase, which can modulate production of isoprenoids and sterols. Perturbation of mevalonate kinase activity from its normal physiological range has been correlated with disease. In particular, mevalonic aciduria results from inherited mevalonate kinase deficiency. The proposed studies will result in the elucidation of structure/function correlations that account for mevalonate kinase catalysis and feedback regulation. This information will be useful for prediction of the consequences of mutations in the human gene that encodes this enzyme. Additionally, such data will facilitate the design of therapeutic strategies aimed at attenuating enzyme activity and potentially diminishing prenylation of cellular targets. Progress toward these long-term objectives will be generated by initial pursuit of four specific aims. (1) Recombinant forms of human and rat mevalonate kinase will be developed and characterized. The purified enzymes will be kinetically and structurally characterized and the mechanism of feedback inhibition established. (2) Using affinity labeling and sited-directed mutagenesis techniques, mevalonate kinase's active site will be identified and the function of active site amino acids assigned. (3) High resolution structures of rat and human mevalonate kinases will be determined by X-ray crystallography. (4) Human mevalonate kinase mutants will be modeled to determine the molecular basis for mevalonic aciduria.

描述：甲瓦硅酸是生物合成的关键中间体 细胞类异戊二烯和固醇。 三种酶，甲谷酸酯激酶， 磷酸综合酸酯激酶和甲氯酸酯5-磷酸脱羧酶是 需要将甲谷酸盐代谢为生物合成活性的异on- 等肾上腺磷酸盐。 最初概述的酶 Mevalonate代谢重点是甲戊酸激酶，可以调节 产生类异丙醇和固醇。 大甲酸激酶的扰动 其正常生理范围的活动与 疾病。 尤其是，甲瓦硅酸尿来自遗传 Mevalonate激酶缺乏。 拟议的研究将导致 阐明结构/功能相关性的甲酸盐 激酶催化和反馈调节。 这些信息将很有用 为了预测人类基因突变的后果 编码此酶。 此外，此类数据将有助于设计 旨在减弱酶活性的治疗策略和潜在的治疗策略 细胞靶标的前介质减少。 朝着这些进展 长期目标将通过对四个特定的初步追求产生 目标。 （1）重组形式的人和大鼠甲位酸酯激酶将是 开发和表征。 纯化的酶将是动力学的，并且 结构表征和反馈抑制的机制 已确立的。 （2）使用亲和力标记并定向诱变 技术，将识别甲瓦龙酸酯激酶的活跃位点，并 分配活性位点氨基酸的功能。 （3）高分辨率 大鼠和人甲龙酸酯激酶的结构将由X射线确定 晶体学。 （4）人类甲丙酸激酶突变体将被建模为 确定甲瓦氏酸尿的分子基础。