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Enzymology of Homoserine Acylation in Methionine Biosynt

蛋氨酸 Biosynt 中高丝氨酸酰化的酶学

基本信息

批准号：
6555934
负责人：
TIMOTHY L BORN
金额：
$ 14.3万
依托单位：
GEORGE MASON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-06-01 至 2006-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6555934
关键词：
active sites acylation acyltransferase aminoacid biosynthesis antibacterial agents crystallization enzyme activity enzyme inhibitors enzyme structure enzyme substrate mass spectrometry methionine protein purification protein structure function site directed mutagenesis

项目摘要

DESCRIPTION (provided by applicant): The methionine biosynthetic pathway, absent in mammals, produces two compounds required for bacterial survival, methionine and S-adenosylmethionine. Disruption of this pathway prevents bacterial growth unless sufficient methionine is obtained from the environment. Therefore, enzymes in this pathway may potentially be targets for novel antibacterial compounds. The first unique step in methionine biosynthesis, acylation of the g-hydroxyl of homoserine, controls flux of homoserine into the pathway. This acylation is catalyzed by one of two enzymes, homoserine transacetylase (HTA) or homoserine transsuccinylase (HTS). The long-term goals of this project are to ascertain whether these enzymes are potential targets for antibacterial agents and to design inhibitors that will function as lead compounds. Initial kinetic characterizations of both enzymes have been reported. In this proposal three specific aims will be pursued. First, the amino acids comprising the Ser-Asp-His catalytic triad of HTA will be identified. This will be accomplished through the combination of sequence alignments, site-directed mutagenesis and steady-state kinetic characterization. Second, the active site residues of HTS, which are different from those of HTA, will be identified. This will be accomplished through a combination of sequence alignment, site-directed mutagenesis, chemical modification, and steady-state kinetic characterization. Third, structural analysis of HTA, HTS, and select mutants will be pursued in an effort to correlate function with structure. All three of these aims directly support the long-term goals of this project. The proposed experiments will be used to train both undergraduate and Master's level students in the area of biochemistry

描述（由申请方提供）：蛋氨酸生物合成途径在哺乳动物中不存在，产生细菌存活所需的两种化合物，蛋氨酸和S-腺苷蛋氨酸。除非从环境中获得足够的甲硫氨酸，否则该途径的破坏会阻止细菌生长。因此，该途径中的酶可能是新型抗菌化合物的潜在靶点。甲硫氨酸生物合成中的第一个独特步骤，高丝氨酸的g-羟基的酰化，控制高丝氨酸进入途径的通量。该酰化由两种酶之一催化，高丝氨酸转乙酰酶（HTA）或高丝氨酸转琥珀酰酶（HTS）。该项目的长期目标是确定这些酶是否是抗菌剂的潜在靶点，并设计将作为先导化合物的抑制剂。这两种酶的动力学特性的初步报告。本建议将追求三个具体目标。首先，将鉴定构成HTA的Ser-Asp-His催化三联体的氨基酸。这将通过序列比对、定点诱变和稳态动力学表征的组合来实现。其次，将鉴定HTS的活性位点残基，其不同于HTA的活性位点残基。这将通过序列比对、定点诱变、化学修饰和稳态动力学表征的组合来实现。第三，HTA，HTS和选择突变体的结构分析将努力将功能与结构相关联。所有这三个目标都直接支持该项目的长期目标。建议的实验将用于培养本科生和硕士生在生物化学领域的水平

项目成果

期刊论文数量（3）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Identification of catalytic cysteine, histidine, and lysine residues in Escherichia coli homoserine transsuccinylase.

DOI：
10.1021/bi0620252
发表时间：
2007-02
期刊：
Biochemistry
影响因子：
2.9
作者：
Katharine Ziegler;Schroeder M Noble;E. Mutumanje;B. Bishop;D. Huddler;T. Born
通讯作者：
Katharine Ziegler;Schroeder M Noble;E. Mutumanje;B. Bishop;D. Huddler;T. Born

Substrate analysis of homoserine acyltransferase from Bacillus cereus.