STRUCTURAL MODELS FOR MOLYBDOENZYMES

钼酶的结构模型

• 批准号: 6045541
• 负责人: JOHN H ENEMARK
• 金额: $ 27.69万
• 依托单位: UNIVERSITY OF ARIZONA
• 依托单位国家: 美国
• 财政年份: 1986
• 资助国家: 美国
• 起止时间: 1986-06-01 至 2003-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6045541
• 关键词: Raman spectrometry; X ray spectrometry; acidity /alkalinity; active sites; circular magnetic dichroism; cofactor; electron nuclear double resonance spectroscopy; electron spin resonance spectroscopy; electron transport; enzyme model; enzyme structure; flash photolysis; heme; iron sulfur protein; metalloenzyme; model design /development; molybdenum; nuclear magnetic resonance spectroscopy; oxidoreductase; physical model; sulfite reductase; sulfites; xanthine oxidase

Molybdenum is an essential trace element for all forms of life, and over 30 molybdoenzymes are known that catalyze oxidation-reduction reactions that are essential in the metabolism of carbon, nitrogen and sulfur. In humans molybdoenzymes play physiologically vital roles in the oxidation of sulfite to sulfate (sulfite oxidase) and in certain aspects of purine metabolism (xanthine oxidase). Fatal simultaneous deficiencies in the activities of both of these enzymes due to an inborn deficiency of a "molybdenum cofactor" have been well documented in children. Point defects in the sulfite oxidase protein itself can also produce the severe neurological symptoms of sulfite oxidase deficiency. These symptoms include dislocated ocular lenses, mental retardation, and, in severe cases, attenuated growth of the brain and early death. Development of methods to clone and express human sulfite oxidase has revealed several different clinical point mutations that result in isolated sulfite oxidase deficiency. The X-ray structure of the highly homologous chicken liver sulfite oxidase provides a molecular basis for interpreting the fatal point mutations of the human enzyme. The research proposed here addresses the fundamental properties of sulfite oxidase by an integrated program of biochemical, biophysical and model compound studies. Emphasis will be given to the use of electron spin echo envelope modulation (ESEEM) spectroscopy to probe the molybdenum environment; to developing pulsed electron-electron double resonance (ELDOR) spectroscopy to probe the molybdenum-iron interaction; and to studying the factors that control the rates of intramolecular electron transfer between the molybdenum and iron centers in native and mutant sulfite oxidase proteins.

钼是所有生命形式所必需的微量元素，已知有30多种氧化还原酶催化碳、氮和硫代谢所必需的氧化还原反应。 在人类中，钼酶在亚硫酸盐氧化为硫酸盐（亚硫酸盐氧化酶）和嘌呤代谢的某些方面（黄嘌呤氧化酶）中发挥着重要的生理作用。 由于先天性缺乏“钼辅因子”，这两种酶的活性同时存在致命性缺陷，这在儿童中已有很好的记录。 亚硫酸氧化酶蛋白本身的点缺陷也可产生亚硫酸氧化酶缺乏症的严重神经症状。 这些症状包括晶状体脱位、智力迟钝，严重时还会导致大脑发育迟缓和早死。 克隆和表达人亚硫酸盐氧化酶的方法的发展揭示了导致孤立的亚硫酸盐氧化酶缺乏症的几种不同的临床点突变。 高度同源的鸡肝亚硫酸盐氧化酶的X射线结构为解释人类酶的致命点突变提供了分子基础。 本研究通过生物化学、生物物理和模型化合物的综合研究，探讨了亚硫酸盐氧化酶的基本性质。重点将给予使用电子自旋回波包络调制（ESEEM）光谱探测钼环境;发展脉冲电子-电子双共振（ELDOR）光谱探测的铼-铁相互作用;和研究的因素，控制钼和铁中心之间的分子内电子转移率在本地和突变亚硫酸氧化酶蛋白质。