STRUCTURE-FUNCTION STUDIES ON ENZYMES INVOLVED IN THE LYSINE BIOSYNTHETIC PATHWA

赖氨酸生物合成途径相关酶的结构功能研究

• 批准号: 8170135
• 负责人: Katherine S Bateman
• 金额: $ 0.03万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170135
• 关键词: Arabidopsis; Bacteria; Chemicals; Chlamydia; Chlamydia Infections; Collaborations; Complex; Computer Retrieval of Information on Scientific Projects Database; Crystallization; Crystallography; Data; Drug Design; Enzymes; Funding; Genus Mycobacterium; Grant; Haemophilus influenzae; Human; Institution; Investigation; Lysine; Lysine Biosynthesis Pathway; Methods; Mouse-ear Cress; Pathway interactions; Plants; Research; Research Personnel; Resolution; Resources; Solutions; Source; Structure; Synchrotrons; Transaminases; Tuberculosis; United States National Institutes of Health; Work; antimicrobial drug; beamline; design; epimerase; high throughput screening; inhibitor/antagonist; mutant; pathogen

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Bacterial biosynthesis of lysine forms an attractive target for the design of new antimicrobial agents because this pathway is indispensable for bacteria and is absent in humans. We have undertaken structural investigations on enzyme targets involved in the biosynthesis of lysine from bacterial pathogens and plants. Crystal structures of two of these enzymes, namely, diaminopimelate epimerase from Haemophilus influenzae and LL-diaminopimelate aminotransferase from Arabidopsis thaliana have been determined recently. Structural work is continuing on these enzymes in complex with inhibitors and of mutant forms to understand details of the catalytic mechanism for the design of effective inhibitors. Recently, we have obtained crystals of the diaminopimelate epimerase from Arabidopsis. Also, crystallization trials are currently underway on the diaminopimelate aminotransferase from the human pathogen, Chlamydia. We have an ongoing collaboration for high-throughput screening of 80,000 chemical compounds in order to find potential inhibitor leads for crystallographic analysis in our drug design efforts against Chlamydial infections. Other targets undergoing crystallization trials include diaminopimelate desuccinylase from the tuberculosis causing mycobacteria. In order to achieve the best possible resolution and quality of diffraction data, as well as for de novo structure solution using MAD/SAD methods, we require access to the synchrotron beamlines for macromolecular crystallography.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 赖氨酸的细菌生物合成成为设计新型抗菌剂的一个有吸引力的目标，因为该途径对于细菌是必不可少的，而在人类中却缺乏。我们对细菌病原体和植物赖氨酸生物合成所涉及的酶靶标进行了结构研究。最近确定了其中两种酶的晶体结构，即来自流感嗜血杆菌的二氨基庚二酸差向异构酶和来自拟南芥的LL-二氨基庚二酸转氨酶。对这些酶与抑制剂复合物和突变形式的结构研究仍在继续，以了解设计有效抑制剂的催化机制的细节。最近，我们从拟南芥中获得了二氨基庚二酸差向异构酶的晶体。此外，目前正在进行针对人类病原体衣原体的二氨基庚二酸转氨酶的结晶试验。我们正在进行合作，对 80,000 种化合物进行高通量筛选，以便在我们针对衣原体感染的药物设计工作中找到用于晶体学分析的潜在抑制剂先导化合物。正在进行结晶试验的其他靶标包括来自引起分枝杆菌的结核病的二氨基庚二酸脱琥珀酰酶。为了实现衍射数据的最佳分辨率和质量，以及使用 MAD/SAD 方法的从头结构解决方案，我们需要使用同步加速器光束线进行大分子晶体学。