Mechanisms of Enzyme Catalysis

描述（由申请人提供）：本项目的目标是确定PEP β/异柠檬酸裂解酶和4-羟基苯甲酰-CoA硫酯酶家族的选定成员的结构、功能和作用机制。这些信息将被用来与活性位点结构的催化，从而确定标记，这可以应用于分配功能的所有未知的蛋白质在每个酶家族。新的蛋白质功能和代谢途径，预计将出现从这些努力，沿着活性位点结构的确定，作为药物发现的基础。最后，从拟议的研究中，首席研究员和她的团队将深入了解介导两个酶家族所代表的不同化学物质的酶的催化机制，以及这些催化机制如何从祖先活性位点模板进化而来。具体目的1-4将阐述PEP β/异柠檬酸裂解酶家族的四个成员的结构、功能和催化机制：洋葱伯克霍尔德氏菌的膦酰丙酮酸水解酶，5，10-亚甲基四氢叶酸：铜绿假单胞菌的3-甲基-2-氧代丁酸羟甲基转移酶，一种与康乃馨花瓣死亡相关的蛋白质，以及一种存在于结核分枝杆菌中的功能未知的蛋白质（Rv 1998 c）。具体目标5-9将阐述4-羟基苯甲酰基-CoA硫酯酶家族的五个成员的结构、功能和催化机制：4-羟基苯甲酰基-CoA硫酯酶，流感嗜血杆菌Tol-pal途径的YgbC酶，大肠杆菌苯乙酸盐分解代谢途径的P76084未知蛋白，耐盐芽孢杆菌上龙胆酸途径的BH 1997未知蛋白和铜绿假单胞菌PA 5519与人长链酰基-CoA硫酯酶的同源物

DESCRIPTION (provided by applicant): The goal of this project is to determine structure, function, and mechanism of action of selected members of the PEP mutase/isocitrate lyase and 4-hydroxybenzoyl-CoA thioesterase enzyme families. This information will be used to relate active site structure to catalysis, and thereby identify markers, which can be applied in the assignment of function to all unknown proteins within each enzyme family. The novel protein functions and metabolic pathways that are anticipated to emerge from these efforts will, along with the active site structure determinations, serve as the foundation for drug discovery. Finally, from the proposed studies the principal investigator and her group will gain insight into the catalytic mechanisms of the enzymes mediating the diverse chemistries represented by the two enzyme families, and into how these catalytic mechanisms evolved from ancestral active site templates. Specific Aims 1-4 will address structure, function and catalytic mechanism in four members of the PEP mutase/ isocitrate lyase enzyme family: phosphonopyruvate hydrolase of Burkholderia cepacia, 5, 1 0-methylenetetrahydrofolate: 3-methyl-2-oxobutanoate hydroxymethyl transferase of Pseudomonas aeruginosa, a protein associated with petal death in carnation, and a protein of unknown function present in Mycobacterium tuberculosis (Rv1998c). Specific Aims 5-9 will address structure, function and catalytic mechanism in five members of the 4-hydroxybenzoyl-CoA thioesterase enzyme family: 4-hydroxybenzoyl-CoA thioesterase, the YgbC enzyme of the Haemophilus influenza Tol-pal Pathway, the P76084 unknown protein of the E.coli Phenylacetate Catabolic Pathway, the BH1 997 unknown protein of the Bacillus halodurans Upper Gentisate Pathway and the Pseudomonas aeruginosa PA551 9 homologue to the Human Long Chain Acyl-CoA Thioesterase