Characterization of new methanobactins and their biosynthetic enzymes

新型甲烷菌素及其生物合成酶的表征

• 批准号: 10228366
• 负责人: Yun Ji Park
• 金额: $ 2.89万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-02-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10228366
• 关键词: Address; Affinity; Anabolism; Bacteria; Binding; Biochemical; Biochemical Genetics; Bioinformatics; Biological; Biological Sciences; Chelating Agents; Chemical Structure; Chemicals; Chemistry; Collaborations; Complex; Copper; Core Protein; Culture Media; Development; Dioxygenases; Educational workshop; Electron Spin Resonance Spectroscopy; Engineering; Ensure; Environment; Enzymes; Equipment; Escherichia coli; Genes; Genetic study; Homeostasis; In Vitro; Interdisciplinary Study; Investigation; Knowledge; Laboratories; Libraries; Ligands; Mass Spectrum Analysis; Metabolic; Metabolic Pathway; Metabolism; Metals; Methane; Methane hydroxylase; Methanol; Methylococcaceae; Methylocystis; Molecular; Molecular Structure; Natural Products; Nitrogen; Operon; Oxazolone; Oxidoreductase; Particulate; Pathway interactions; Peptides; Photons; Preparation; Proteins; Research; Resources; Ribosomes; Role; Source; Specificity; Structure; Sulfur; System; Techniques; Therapeutic; Thioamides; Training; Training Programs; Transaminases; Universities; Vertebral column; X-Ray Crystallography; amino group; beamline; biophysical techniques; career; chemical property; cofactor; drug candidate; education research; enzyme biosynthesis; genetic approach; genetic manipulation; human disease; in vivo; insight; metal chelator; novel; novel therapeutics; oxidation; physical property; programs; protein expression; success; sulfotransferase; training opportunity; transamination

Project Summary/Abstract Methanobactins (Mbns) are ribosomally-synthesized and post-translationally modified peptide natural products produced by methanotrophic (methane-oxidizing) bacteria to obtain copper from the environment; copper is necessary for their primary metabolic enzyme, copper- dependent particulate methane monooxygenase (pMMO). Previously characterized Mbns contain a peptidic backbone with two heterocycle/thioamide pairs that provide two nitrogen and two sulfur ligands for Cu(I). These Mbns bind copper with high affinity and specificity, and are under investigation as drug candidates for human diseases of copper metabolism. Recent bioinformatics studies have revealed that there are new types of Mbns that are predicted to differ in structure and chemical properties from previously characterized Mbns. Moreover, several uncharacterized enzymes that are expected to be involved in the biosynthesis of these new Mbns are not well understood. In this project, uncharacterized Mbns with predicted novel structures will be explored, and the enzymes responsible for their biosynthesis will be studied by both in vivo and in vitro approaches. This research will provide significant insights into the diversity of Mbn structures and functions, will elucidate new enzymatic chemistries, and will impact development of Mbns and Mbn-like molecules as therapeutics. A multifaceted training program has been planned. Training in a variety of biochemical and genetic as well as spectroscopic/spectrometric techniques needed to isolate and characterize Mbns and their biosynthetic enzymes will be provided in the Rosenzweig laboratory. In addition, formal training opportunities outside the laboratory, such as programs at the Life Sciences Collaborative Access Team (LS-CAT) beamlines at the Advanced Photon Source (APS) at Argonne National Laboratory and at the Penn State University Bioinorganic Training Workshop, both of which are closely related to the proposed research, will be available. The Department of Molecular Biosciences at Northwestern University is equipped with all the equipment and resources necessary to conduct this research. In particular, collaborations with the Hoffman laboratory for electron paramagnetic resonance (EPR) spectroscopy, Dr. Zhang at the Integrated Molecular Structure Education and Research Center (IMSERC) for a suite of NMR spectroscopic techniques, and Kelleher group for mass spectrometry will facilitate carrying out this research. These training opportunities combined with the collaborative and scientific environment at Northwestern University will ensure the success of the proposed research and will provide excellent preparation for an independent, interdisciplinary research career.

项目概要/摘要 甲烷菌素 (Mbns) 由核糖体合成并经过翻译后修饰 由甲烷氧化（甲烷氧化）细菌产生的肽天然产物以获得 环境中的铜；铜是它们的主要代谢酶所必需的，铜- 依赖颗粒甲烷单加氧酶 (pMMO)。先前表征的 Mbns 包含具有两个杂环/硫代酰胺对的肽主链，提供两个氮和 Cu(I) 的两个硫配体。这些 Mbns 以高亲和力和特异性结合铜，并且 正在研究作为人类铜代谢疾病的候选药物。最近的 生物信息学研究表明，预计存在新型 Mbns 结构和化学性质与之前表征的 Mbns 不同。而且， 一些未表征的酶预计参与这些物质的生物合成 新的Mbns还没有被很好地理解。在这个项目中，未表征的 Mbns 与预测的小说 将探索结构，并研究负责其生物合成的酶 体内和体外方法。这项研究将为以下方面提供重要见解： Mbn 结构和功能的多样性，将阐明新的酶化学，并将 影响 Mbns 和 Mbn 类分子作为治疗药物的发展。 已规划了多方面的培训计划。各种生化培训 以及分离和分离所需的遗传以及光谱/光谱技术 Rosenzweig 实验室将提供表征 Mbns 及其生物合成酶。 此外，实验室外的正式培训机会，例如 Life 项目 先进光子源的科学协作访问团队 (LS-CAT) 光束线 （APS）在阿贡国家实验室和宾夕法尼亚州立大学生物无机培训 研讨会将举行，两者都与拟议的研究密切相关。这 西北大学分子生物科学系装备齐全 进行这项研究所需的设备和资源。特别是与以下机构的合作： 霍夫曼电子顺磁共振 (EPR) 光谱实验室，张博士 综合分子结构教育和研究中心（IMSERC）的一套 核磁共振波谱技术和凯莱赫质谱分析小组将有助于携带 出这项研究。这些培训机会与协作和科学相结合 西北大学的环境将确保拟议研究的成功 将为独立的跨学科研究生涯提供良好的准备。