STRUCTURE AND MECHANISM OF HOTDOG-FOD THIOESTERASES

热狗食品硫酯酶的结构和机制

• 批准号: 8363404
• 负责人: Karen A Allen
• 金额: $ 0.23万
• 依托单位: BROOKHAVEN SCIENCE ASSOC-BROOKHAVEN LAB
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363404
• 关键词: Acyl Carrier Protein; Acyl Coenzyme A; Aromatic Compounds; Binding; Biochemical; Biochemistry; Carboxylic Acids; Catalysis; Cell physiology; Coenzyme A; Crystallography; Elements; Enzymes; Escherichia coli; Family; Family member; Fatty Acids; Funding; Goals; Grant; Hereditary Disease; Human; Human Genetics; Hydrolysis; Knowledge; Life; Light; Lipids; Malignant neoplasm of brain; Metabolic; National Center for Research Resources; Obesity; Organism; Parents; Play; Principal Investigator; Pseudomonas aeruginosa; Reaction; Research; Research Infrastructure; Resources; Role; Source; Structure; Sulfhydryl Compounds; Synchrotrons; Transcriptional Regulation; United States National Institutes of Health; Virulence; beta pleated sheet; chemical reaction; comparative; cost; design; fatty acid biosynthesis; inhibitor/antagonist; lipid metabolism; pathogen; phenylacetic acid; thioester

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. Thioesters are ubiquitous metabolites and regulators, especially relevant, but not limited, to lipid metabolism. Thioesters are removed through the action of thioesterases, which catalyze hydrolysis reactions to generate parent carboxylic acids and thiols. A majority of these enzymes belong to the hotdog thioesterase family targeted in our studies. The hotdog thioesterase family functions to regulate the levels of acyl-CoA metabolites and acyl-Acyl Carrier Proteins (ACPs) in all forms of life. A wide variety of hotdog thioesterases function within each organism (viz, 21 in humans; 29 in Pseudomonas aeruginosa). The hotdog thioesterases were first characterized by biochemists as type II thioesterases, functioning in fatty acid and polyketide synthesis. Despite the fact that they catalyze common chemical reactions, the hotdog fold thioesterases display a wide range of catalytic motifs and substrate recognition elements. Although this diversity provides a targeting advantage for specific inhibitors, it demands the identification, comparative structure analysis, and determination of function and catalytic mechanism for a variety of family members. Our plan is to carry out a detailed study of selected thioesterases in order to elucidate the underlying principles that govern substrate recognition and catalysis, develop a strategy for the design of specific inhibitors, and expand our knowledge of thioester biochemistry. The goal of the study is to determine biochemical functions, catalytic mechanisms and tight binding inhibitors for hotdog thioesterases that are implicated in lipid-related, human genetic disorders that underlie obesity and brain cancer, and that are required for virulence of bacterial pathogens for humans. Our immediate targets include the five E. coli hotdog thioesterases, YbdB, YbcG, YbaW, YicA, Ycil, which are also found in a range of human pathogens. (funded by NIH GM028688).This Hotdog domain is involved in many cellular processes which range from thiester hydrolysis, transcriptional regulation of fatty acid biosynthesis, to the degredation of aromatic compounds such as phenylacetic acid. The "Hotdog fold" consists of seven stranded antiparallel beta-sheets as the "bun", which wraps around a five-turn alpha-helical "sausage". By studying this domain within the thioesterase superfamily we hope to understand the metabolic role it plays as a ubiquitous motif found in all branches of life. In particular we are studying BH1999 thioesterase, 3-HBA CoA thioesterase, and PaaI thioesterase.

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 硫酯是普遍存在的代谢物和调节剂，尤其与脂质代谢相关，但不限于此。通过硫酯酶的作用除去硫酯，硫酯酶催化水解反应以产生母体羧酸和硫醇。这些酶中的大多数属于我们研究中靶向的热狗硫酯酶家族。 热狗硫酯酶家族的功能是调节所有生命形式中酰基-CoA代谢物和酰基-酰基载体蛋白（ACP）的水平。各种各样的热狗硫酯酶在每种生物体中发挥作用（即，人类中有21种;铜绿假单胞菌中有29种）。热狗硫酯酶首先被生物化学家表征为II型硫酯酶，在脂肪酸和聚酮化合物合成中起作用。尽管它们催化常见的化学反应，热狗折叠硫酯酶显示出广泛的催化基序和底物识别元件。虽然这种多样性为特异性抑制剂提供了靶向优势，但它需要对各种家族成员进行鉴定、比较结构分析以及确定功能和催化机制。 我们的计划是进行一个详细的研究，选择硫酯酶，以阐明底物识别和催化的基本原则，制定一项战略，设计特定的抑制剂，并扩大我们的知识硫酯生物化学。这项研究的目的是确定热狗硫酯酶的生化功能，催化机制和紧密结合抑制剂，这些酶与肥胖和脑癌的脂质相关的人类遗传疾病有关，并且是人类细菌病原体毒力所必需的。我们的直接目标包括五个E。大肠杆菌热狗硫酯酶，YbdB、YbcG、YbaW、YicA、YcB，其也在一系列人类病原体中发现。该Hotdog结构域涉及许多细胞过程，其范围从硫酯水解、脂肪酸生物合成的转录调节到芳族化合物如苯乙酸的降解。“热狗折叠”由七个反平行的β折叠组成，作为“面包”，它包裹着一个五圈的α螺旋“香肠”。通过研究硫酯酶超家族中的这个结构域，我们希望了解它作为一个在生命的所有分支中发现的普遍存在的基序所起的代谢作用。特别地，我们正在研究BH 1999硫酯酶、3-HBA CoA硫酯酶和Pastine硫酯酶。