Structural Models For Molybdoenzymes

钼酶的结构模型

• 批准号: 7540368
• 负责人: JOHN H ENEMARK
• 金额: $ 37.43万
• 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 2011-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7540368
• 关键词: Active Sites; Address; Attenuated; Benchmarking; Biological Process; Brain; Cell Nucleus; Cessation of life; Chemicals; Child; Circular Dichroism Spectroscopy; Classification; Crystalline Lens; Defect; Deuterium; Electron Nuclear Double Resonance; Electron Spin Resonance Spectroscopy; Electron Transport; Electronics; Electrons; Enzymes; Frequencies; Gases; Growth; Health; Heme; Human; Iron; Kinetics; Label; Lasers; Libraries; Ligands; Magnetism; Measurement; Mental Retardation; Metals; Molecular; Molecular Structure; Molybdenum; Mus; Mutation; Neurologic; Nuclear; Oxidoreductase; Pathway interactions; Phase; Physiologic pulse; Physiological; Property; Proteins; Raman Spectrum Analysis; Recombinants; Research; Retinaldehyde; Roentgen Rays; Role; Spectrum Analysis; Structural Models; Structure; Sulfites; Symptoms; Techniques; Theoretical model; Transition Elements; Tungsten; Water; X-Ray Crystallography; Xanthines; biochemical model; circular magnetic dichroism; clinically significant; electron nuclear double resonance spectroscopy; electronic structure; enzyme activity; flash photolysis; molybdenum cofactor; mutant; professor; programs; protein structure; sulfite dehydrogenase; sulfite oxidase

DESCRIPTION (provided by applicant): Molybdenum and tungsten are the only second and third row transition elements that have known biological functions. At present, over 50 molybdenum enzymes are known, and several are of clinical significance (xanthine and retinaldehyde oxidoreductases, sulfite oxidase) for human health. Fatal simultaneous deficiencies in the activities of these enzymes due to an inborn deficiency of a "molybdenum cofactor" have been well documented in children. Point defects in the sulfite oxidase (SO) protein can also produce the 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. The research proposed here addresses the fundamental properties of sulfite oxidase and other molybdoenzymes by an integrated program of biophysical, biochemical and model compound studies. Variable frequency pulsed electron paramagnetic resonance (EPR) techniques, especially electron spin echo envelope modulation (ESEEM), pulsed electron-nuclear double resonance (ENDOR), and electron-electron double resonance (ELDOR) spectroscopies, will be used to probe in detail the surroundings of the molybdenum active site in wild-type and pathogenic mutants of SO through labeling with deuterium, 17O and 33S, and by complementary theoretical calculations. Recombinant mouse SO, which is highly homologous to human SO, will be used to generate a library of physiological and non- physiological mutants of mammalian SO for systematic structural, chemical, mechanistic and spectroscopic studies. The rates of intramolecular electron transfer between the molybdenum and iron centers of SO will be investigated by flash photolysis and theoretical modeling.

描述（由申请人提供）：钼和钨是唯一具有已知生物功能的第二和第三行过渡元素。目前已知的钼酶有50多种，其中几种对人体健康具有临床意义（黄嘌呤和视黄醛氧化还原酶、亚硫酸盐氧化酶）。在儿童中，由于先天缺乏“钼辅助因子”，这些酶的活性同时存在致命的缺陷。亚硫酸盐氧化酶（SO）蛋白的点缺陷也可引起亚硫酸盐氧化酶缺乏症的神经症状。这些症状包括晶状体脱位、智力迟钝，在严重的情况下，还会导致大脑发育迟缓和过早死亡。本文通过生物物理、生化和模型化合物的综合研究，探讨了亚硫酸盐氧化酶和其他钼酸酶的基本性质。利用变频脉冲电子顺磁共振（EPR）技术，特别是电子自旋回波包络调制（ESEEM）、脉冲电子-核双共振（ENDOR）和电子-电子双共振（ELDOR）光谱，通过氘、17O和33S标记，并通过互补的理论计算，详细探测SO野生型和致病性突变体钼活性位点周围环境。重组小鼠SO与人类SO高度同源，将用于建立哺乳动物SO生理和非生理突变文库，用于系统的结构、化学、机制和光谱研究。利用闪光分解和理论建模的方法研究了钼和铁中心之间的分子内电子转移速率。