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.

抽象的 金属离子与蛋白质的结合提供了独特的化学反应性，这是许多化学反应的核心 我的实验室探究了自然界最令人惊奇的化学转化。 超亲核试剂和自由基辅助因子的反应性，同时保护自身免受潜在损害。 对于生物信息学和基因组研究来说，这是一个令人难以置信的激动人心的时刻。 以令人眼花缭乱的速度识别新的假定金属酶，目前有超过 100,000 个独特序列 与自由基 S-腺苷甲硫氨酸 (SAM) 酶家族相关。 令人印象深刻的是，酶正在揭示化学和新的辅因子结合结构基序。 这些 Radical SAM (RS) 酶是生产天然产物的生物合成途径的一部分 新颖的分子支架和有前途的药物特性（包括抗生素、抗病毒和抗- 我的实验室正在采用我们最喜欢的 X 射线晶体学技术来探测。 该家族内的序列空间，目的是了解 RS 酶如何利用自由基物种 在接下来的五年中，我们将利用最近的成功并继续进行具有挑战性的化学反应。 研究钴胺素依赖性 RS 酶的结构/功能，该酶有 7000 个成员。 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 最近备受关注的不仅是 RS 酶家族，还有超亲核试剂反应性。 甘氨酰自由基酶 (GRE) 家族也受到越来越多的关注。 微生物组项目正在提供有关 GRE 在人类中的重要性和丰度的新信息 例如，肠道中发现的最丰富的未表征酶是 GRE！ 未来五年，我们计划调查 GRE 家族中几个新发现的成员，这些成员似乎 成为人类微生物群落的关键参与者。我们的目标是使用我们的结构工具来探究。 有助于微生物代谢的自由基化学的分子基础，并可能 在人类肠道中，许多 GRE 都存在于常见病原体中，例如艰难梭菌、 最后，现在是研究“生命大簇”的好时机，它们是 负责固定碳（C 簇/A 簇）、氮（MoFe 簇）和氢（H 簇）。 我的实验室专注于碳固定以及一氧化碳脱氢酶/乙酰基的 C 和 A 簇 CoA 合酶的最新进展提供了重组系统，使我们能够探测簇。 以以前不可能的方式研究组装、反应机制和氧敏感性。 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来 研究 C 簇氧敏感性的分子基础。