Mechanisms of Binuclear Non-Heme Iron Enzymes

双核非血红素铁酶的机制

• 批准号: 6740052
• 负责人: MICHAEL D CLAY
• 金额: $ 4.3万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-02-01 至 2006-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6740052
• 关键词: Raman spectrometry; active sites; chemical structure; computational biology; electron spin resonance spectroscopy; enzyme mechanism; enzyme structure; ferritin; infrared spectrometry; iron; postdoctoral investigator; structural biology

DESCRIPTION (provided by applicant): Binuclear non-heme iron active sites are present in a wide variety of proteins and enzymes which perform different biological functions. This proposal will specifically investigate those sites present in sMMO, R2, D9D and frog M-ferritin to elucidate their differences in dioxygen reactivity (i.e. hydroxylation, radical generation, desaturation and ferroxidation). These differences have led to the identification of various diiron peroxy (P) and high-valent (Q and X) intermediates. While these enzymatic intermediates have been investigated, relatively little is known about their geometric and electronic structure and, more importantly, how their structure relates to reactivity. Therefore, the major objectives of this research proposal are to use biophysical spectroscopic techniques (EPR, resonance Raman and UV-vis/Near IR absorption/CDNTVH MCD) to characterize the geometric, electronic and magnetic properties of the various diiron peroxy (P) and high-valent (Q and X) intermediates in these enzymes and relevant model complexes and apply computational methodologies to gain greater insight into the mechanistic conversion between these intermediates.

描述（由申请人提供）：双核非血红素铁活性位点存在于执行不同生物功能的多种蛋白质和酶中。该提案将专门研究 sMMO、R2、D9D 和青蛙 M-铁蛋白中存在的位点，以阐明它们在双氧反应性（即羟基化、自由基生成、去饱和和亚铁氧化）方面的差异。这些差异导致了各种过氧化二铁 (P) 和高价（Q 和 X）中间体的鉴定。虽然已经对这些酶中间体进行了研究，但人们对它们的几何和电子结构以及更重要的是它们的结构与反应性的关系知之甚少。因此，本研究计划的主要目标是使用生物物理光谱技术（EPR、共振拉曼和紫外可见/近红外吸收/CDNTVH MCD）来表征各种过氧化二铁（P）和这些酶和相关模型复合物中的高价（Q 和 X）中间体，并应用计算方法来更深入地了解这些中间体之间的机械转化。