Metalloenzyme structure, function and assembly

金属酶的结构、功能和组装

• 批准号: 10413652
• 负责人: CATHERINE L DRENNAN
• 金额: $ 5.55万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2023-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10413652
• 关键词: Acetyl Coenzyme A; Amaze; Antibiotic Resistance; Antibiotics; Antineoplastic Agents; Antiviral Agents; Attention; Binding; Biochemistry; Bioinformatics; Biotechnology; Carbon monoxide dehydrogenase; Chemicals; Chemistry; Clostridium difficile; Cobalamin; Complex; Drug Targeting; Enzymes; Family; Five-Year Plans; Genomics; Goals; Health; Heart; Human; Human Microbiome; Hydrogen; Ions; Laboratories; Life; Metabolism; Metalloproteins; Metals; Molecular; Natural Products; Nature; Nitrogen; Oral cavity; Oxygen; Pathogenesis; Pathway interactions; Pharmacologic Substance; Production; Property; Proteins; Reaction; Recombinants; S-Adenosylmethionine; Structure; Subgroup; System; Techniques; Time; Toxic Environmental Substances; X-Ray Crystallography; base; carbon fixation; climate change; cofactor; frontier; human microbiota; insight; member; metalloenzyme; microbial; novel; pathogen; remediation; scaffold; success; tool; tumor

Abstract The combination of metal ions with proteins offers unique chemical reactivities, which are at the heart of many of Nature's most amazing chemical transformations. My laboratory interrogates how metalloenzymes harness the reactivity of supernucleophiles and radical cofactors, while protecting themselves from potential damage. It is an incredibly exciting time to be studying metalloenzymes. Bioinformatics and genomic studies are identifying new putative metalloenzymes at a dizzying pace, with more than 100,000 unique sequences now associated with the Radical S-adenosylmethionine (SAM) enzyme family alone. Characterization of these enzymes is revealing unprecedented chemistry and new cofactor-binding structural motifs. Impressively, many of these Radical SAM (RS) enzymes are part of biosynthetic pathways that produce natural products with novel molecular scaffolds and promising pharmaceutical properties (including antibiotic, antiviral, and anti- tumor properties). My laboratory is employing our favorite technique of X-ray crystallography to probe sequence space within this family with the goal of understanding how RS enzymes harness radical-species to perform chemically challenging reactions. In the next five years, we will leverage recent success and continue to investigate the structure/function of cobalamin-dependent RS enzymes. This 7000-membered RS subgroup represents a new set of challenges and opportunities to understand how Nature tunes and controls both radical and supernucleophile reactivities. It is not only the RS enzyme family that has been in the spotlight recently; the glycyl radical enzyme (GRE) family is also receiving increased attention. In this latter case, the human microbiome project is providing new information as to the importance and abundance of GREs in the human gut and oral cavities. For example, the most abundant uncharacterized enzyme found in the gut is a GRE! In the next five years, we plan to investigate several newly discovered members of the GRE family that appear to be key players in human microbial communities. Our goal is to use our structural tools to interrogate the molecular basis for the radical-based chemistry that contributes to microbial metabolism, and potentially pathogenesis, in the human gut. A number of these GREs are found in common pathogens, like C. difficile, and are potential drug targets. Finally, it is a great period to be working on the “great clusters of life,” which are responsible for the fixation of carbon (C-cluster/A-cluster), nitrogen (MoFe cluster) and hydrogen (H-cluster). My laboratory focuses on carbon fixation and the C- and A-clusters of carbon monoxide dehydrogenase/acetyl- CoA synthase. Recent advances have afforded recombinant systems that are allowing us to probe cluster assembly, reaction mechanism, and oxygen-sensitivity in a manner that was not possible previously. Oxygen- sensitivity is the Achilles heel of a complex metalloprotein and we plan to use our structural toolbox to investigate the molecular basis of C-cluster oxygen-sensitivity.

摘要 金属离子与蛋白质的结合提供了独特的化学反应性，这是许多蛋白质的核心。 自然界最惊人的化学变化。我的实验室研究金属酶如何利用 超亲核试剂和自由基辅因子的反应性，同时保护自身免受潜在的损害。它 是研究金属酶的一个令人难以置信的激动时刻。生物信息学和基因组研究是 以令人眼花缭乱的速度鉴定新的假定金属酶，现在有超过10万个独特的序列， 与自由基S-腺苷甲硫氨酸（SAM）酶家族单独相关。表征这些 酶揭示了前所未有的化学和新的辅因子结合结构基序。令人印象深刻的是，许多 这些自由基SAM（RS）酶是生物合成途径的一部分， 新的分子支架和有前途的药物特性（包括抗生素、抗病毒和抗- 肿瘤特性）。我的实验室正在使用我们最喜欢的X射线晶体学技术来探测 序列空间在这个家庭的目标是了解RS酶如何利用自由基物种， 进行化学挑战性反应。在未来五年，我们将利用最近的成功，继续 研究钴胺素依赖性RS酶的结构/功能。这个7000元RS子群 代表了一系列新的挑战和机遇，以了解自然如何调整和控制激进的 和超亲核反应性。最近备受关注的不仅仅是RS酶家族; 甘氨酰自由基酶（GRE）家族也受到越来越多的关注。在后一种情况下， 微生物组项目提供了关于GRES在人类中的重要性和丰富性的新信息。 肠道和口腔。例如，在肠道中发现的最丰富的未表征的酶是GRE！在 在接下来的五年里，我们计划研究几个新发现的GRE家族成员，这些成员似乎 成为人类微生物群落的关键角色。我们的目标是使用我们的结构工具来询问 分子基础的自由基为基础的化学，有助于微生物代谢，并可能 致病机制，在人类肠道中。这些GREs中的许多存在于常见的病原体中，如C。difficile， 并且是潜在的药物靶点。最后，这是一个伟大的时期，致力于“伟大的生命集群”， 负责碳（C-簇/A-簇）、氮（MoFe簇）和氢（H-簇）的固定。 我的实验室专注于碳固定和一氧化碳脱氢酶/乙酰基- 辅酶A合成酶。最近的进展提供了重组系统，使我们能够探测集群 组装、反应机制和氧敏感性，这在以前是不可能的。氧气- 敏感性是复杂金属蛋白的致命弱点，我们计划使用我们的结构工具箱， 探讨了C团簇氧敏性的分子基础。