Crystallographic Analysis of GHMP kinases

GHMP 激酶的晶体学分析

• 批准号: 6526195
• 负责人: HONG ZHANG
• 金额: $ 21.02万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-08-07 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6526195
• 关键词: X ray crystallography; alcohol phosphotransferase; bacteria; bacterial proteins; biochemical evolution; computer simulation; conformation; crystallization; enzyme inhibitors; enzyme substrate analog; enzyme substrate complex; fungal proteins; intermolecular interaction; magnesium ion; model design /development; molecular cloning; molecular dynamics; molecular site; phosphorylation; physical model; protein folding; protein purification; protein structure function; proteomics; structural biology; yeasts

DESCRIPTION: (provided by applicant) The GHMP class of small metabolite kinases participates in several essential metabolic processes, such as glycolysis, amino acid biosynthesis, and sterol biosynthesis. Currently, the GHMP superfamily contains more than 170 proteins, representing about 12-13 different functions, including galactokinases, homoserine kinases, mevalonate kinases, phosphomevalonate kinases (hence GHMP), mevalonate diphosphate decarboxylase, isopentenyl monophosphate kinase, and archaeal shikimate kinase. Deficiencies in GHMP enzyme activities cause auxotrophic phenotypes in bacteria and hereditary metabolic diseases in humans. Structural characterization of GHMP enzymes and their complexes with substrates is crucial for understanding the active site, catalytic mechanism, inhibition and regulation of these enzymes. The first three dimensional structure of a GHMP protein, the homoserine kinase (HK), revealed a novel nucleotide-binding fold and a unique ATP binding mode. The crystals of the ternary complex between HK and its substrates were also obtained. Structural analysis of the HK-substrate complexes will reveal the key catalytic residues in the phosphoryl transfer reaction and the residues responsible for the substrate specificity. To learn how GHMP-fold accommodates substrates of very different structures, other members in the GHMP superfamily are selected for the structural studies in a systematic approach. Diffracting crystals of the archaeal shikimate kinase have been obtained. Comparative analysis of the enzyme-substrate complexes from different members of the GHMP superfamily will reveal the structural determinants of the substrate specificity in each family, provide a foundation for the structure-base drug design, and further our understanding of the structure-function evolution of this important class of enzymes.

描述：（由申请人提供）GHMP 类小代谢激酶 参与几个重要的代谢过程，例如糖酵解， 氨基酸生物合成和甾醇生物合成。目前，GHMP 超家族包含 170 多种蛋白质，代表约 12-13 种不同的蛋白质 功能，包括半乳激酶、高丝氨酸激酶、甲羟戊酸激酶、 磷酸甲羟戊酸激酶（因此 GHMP）、甲羟戊酸二磷酸脱羧酶、 异戊烯单磷酸激酶和古菌莽草酸激酶。不足之处 GHMP 酶活性导致细菌营养缺陷型 人类遗传性代谢疾病。 GHMP 的结构表征 酶及其与底物的复合物对于理解酶及其与底物的复合物至关重要 这些酶的活性位点、催化机制、抑制和调节。 GHMP 蛋白的第一个三维结构，即高丝氨酸激酶 (HK)，揭示了一种新的核苷酸结合折叠和独特的 ATP 结合模式。 HK 与其基底之间的三元配合物的晶体也被 获得。香港基底复合体的结构分析将揭示关键 磷酰基转移反应中的催化残基和残基 负责底物特异性。了解 GHMP-fold 如何适应 结构非常不同的底物，GHMP 超家族的其他成员 以系统的方法选择进行结构研究。衍射 已获得古菌莽草酸激酶的晶体。比较 GHMP 不同成员的酶-底物复合物分析 超家族将揭示底物的结构决定因素 每个家族的特异性，为结构基础药物提供基础 设计，并进一步我们对结构功能演变的理解 这一类重要的酶。