RESONANCE SPECTROSCOPY OF METALLOPROTEINS

金属蛋白的共振光谱

• 批准号: 3268918
• 负责人: PETER G DEBRUNNER
• 金额: $ 25.13万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 1978
• 资助国家: 美国
• 起止时间: 1978-01-01 至 1994-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3268918
• 关键词: Basidiomycetes; Mossbauer spectrometry; Pseudomonas; acid phosphatase; bacterial proteins; conformation; cytochrome P450; cytochrome oxidase; electron nuclear double resonance spectroscopy; electron spin resonance spectroscopy; electron transport; electronic spectra; enzyme complex; enzyme mechanism; enzyme structure; enzyme substrate; free radical oxygen; hemerythrin; hemoprotein structure; lignin; metalloproteins; molecular energy level; myoglobin; oxidation reduction reaction; peroxidases; protein structure function

The aim of the proposed research is to elucidate the electronic structures of metallo-protein active centers using primarily Mossbauer, EPR, and ENDOR spectroscopy. Of particular interest are the heme proteins that interact with O2 or H2O2 and their various intermediates. The ultimate goal is to understand the structural and dynamic features that control reactivity and function. The following systems will be studied: (i) Work in progress on O2 activation and product formation in the bacterial monoxygenase cytochrome P450cam will continue. Earlier data suggest that an electron can be transferred to the ternary complex of cytochrome P450cam with substrate and O2 by low-temperature X-ray irradiation, and that new EPR- active heme species are formed on stepwise annealing to higher temperatures. The nature of these intermediates will be explored and the products of the reaction will be analyzed. Low-temperature techniques will also be used to search for intermediates in the product-forming steps of the native system and with modified substrates and proteins. (ii) Prof. Sligar's group has produced genetically modified myoglobins, which will be characterized by Mossbauer, EPR and ENDOR spectroscopy in a collaborative effort to correlate amino acid substitutions with changes in function, structure and electronic state. (iii) A Mossbauer study of lignin and manganese peroxidase from the white rot fungus Phanerochaete chrysosporium and of their reaction products with H2O2 and substrate will be started in collaboration with Prof. Gold, Oregon Graduate Institute. (iv) Two terminal oxidases, cytochrome o and cytochrome d from E. coli, will be investigated by EPR, ENDOR and Mossbauer spectroscopy in collaboration with Prof. Gennis. (v) Work on hemerythrin and structurally related binuclear iron proteins and model compounds will continue.

研究的目的是阐明电子结构 金属蛋白质活性中心主要使用穆斯堡尔，EPR和ENDOR 谱 特别令人感兴趣的是血红素蛋白， 与O2或H2 O2及其各种中间体反应。 最终目标是 了解控制反应性的结构和动力学特征， 功能 将研究下列系统： 细菌单加氧酶中O2的活化和产物形成 细胞色素P450 cam将继续。 早期的数据表明， 可以转移到细胞色素P450 cam与 衬底和O2低温X射线辐照，和新的EPR- 活性血红素物种在逐步退火到更高温度时形成。 温度 将探讨这些中间体的性质， 将分析反应产物。 低温技术将 也可用于搜索产品形成步骤中的中间体， 天然系统和修饰的底物和蛋白质。 (ii)教授 Sligar的研究小组已经生产出转基因肌红蛋白， 其特征在于穆斯堡尔，EPR和ENDOR光谱在合作 努力将氨基酸取代与功能变化相关联， 结构和电子态。 (iii)木质素和纤维素的穆斯堡尔谱研究 白色腐真菌黄孢原毛革菌锰过氧化物酶 以及它们与H2 O2和底物的反应产物将在 与俄勒冈州研究生院的Gold教授合作。 (iv)两 末端氧化酶、细胞色素o和细胞色素d。大肠杆菌， 通过EPR，ENDOR和穆斯堡尔光谱研究， 教授杰尼斯 (v)关于血红蛋白和结构上相关的双核 铁蛋白和模型化合物将继续。