Rapid Scanning Stopped Flow Spectrophotometer for Enzymology

用于酶学的快速扫描停流分光光度计

• 批准号: 9604702
• 负责人: Gary Cecchini
• 金额: --
• 依托单位: University of California-San Francisco
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-02-15 至 1998-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9604702&HistoricalAwards=false
• 关键词: Rapid; Scanning; Stopped; Flow; Spectrophotometer

Mechanistic and kinetic studies of enzymes involved in the metabolism of aromatic hydrocarbons, degradation of alkylamines, and respiration will be undertaken. The enzymes include the flavocytochrome p-cresol methylhydroxylase (Pseudomonas putida), the quinoprotein methylamine dehydrogenase (methylotrophic bacteria), copper-containing quinoprotein amine oxidases (human and Arthrobacter globoformis), and iron-sulfur flavoproteins succinate dehydrogenase (mammalian and bacteria) and fumarate reductase from Escherichia coli. The numbers, types, and redox potentials of the organic and metal ion cofactors are already defined. The majority of these enzymes have been cloned to allow for alteration of their properties by genetic manipulation. The available x-ray structures of many of the proteins studied will guide some of the research. It is proposed that the requested rapid-scanning stopped-flow spectrophotometer be used to define events occurring on substrate/ligand binding to enzymes, and the properties of the individual cofactors in integrated functions. It is also proposed that the instrument be used as a rapid-recording, dual-wavelength spectrophotometer for essential quinol oxidation reduction assays.

参与芳香烃代谢、烷基胺降解和呼吸作用的酶的机理和动力学研究将进行。这些酶包括黄细胞色素对甲酚甲基羟化酶（恶臭假单胞菌）、醌蛋白甲胺脱氢酶（甲基营养性细菌）、含铜醌蛋白胺氧化酶（人类和球状节杆菌）、铁硫黄蛋白琥珀酸脱氢酶（哺乳动物和细菌）和大肠杆菌的富马酸还原酶。有机和金属离子辅因子的数量、类型和氧化还原电位已经定义。这些酶中的大多数已经被克隆，允许通过基因操作来改变它们的特性。所研究的许多蛋白质的可用x射线结构将指导一些研究。建议使用要求的快速扫描止流分光光度计来定义底物/配体与酶结合时发生的事件，以及综合功能中单个辅因子的性质。还建议将该仪器作为快速记录双波长分光光度计用于必要的喹啉氧化还原测定。