Characterization of new methanobactins and their biosynthetic enzymes

项目总结/摘要 甲烷氧化菌素（Mbns）是核糖体合成和后修饰的 由甲烷营养（甲烷氧化）细菌产生的肽天然产物， 铜从环境中;铜是必要的，他们的主要代谢酶，铜- 依赖性颗粒甲烷单加氧酶（pMMO）。先前表征的Mbns 含有具有两个杂环/硫代酰胺对的肽骨架，所述杂环/硫代酰胺对提供两个氮， Cu（I）的两个硫配体。这些Mbn以高亲和力和特异性结合铜，并且是 作为治疗人类铜代谢疾病的候选药物正在研究中。最近 生物信息学研究表明，有新类型的Mbns被预测为 在结构和化学性质上与先前表征的Mbns不同。此外，委员会认为， 几种未知的酶，预计参与这些生物合成 新的MBN还没有被很好地理解。在这个项目中，未表征的Mbns与预测的新的 结构将被探索，负责其生物合成的酶将被研究， 包括体内和体外方法。这项研究将提供重要的见解， MBN结构和功能的多样性，将阐明新的酶化学， 影响Mbn和Mbn样分子作为治疗剂的发展。 一个多方面的培训计划已经制定。各种生化训练 和遗传以及光谱/光谱技术需要分离和 将在Rosenzweig实验室提供表征Mbns及其生物合成酶的材料。 此外，实验室外的正式培训机会，如生命科学院的项目， 科学协作访问团队（LS-CAT）在先进光子源的光束线 (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，这两个都是密切相关的拟议研究，将提供。的 西北大学分子生物科学系配备了所有 进行这项研究所需的设备和资源。特别是，与 霍夫曼实验室电子顺磁共振（EPR）光谱，张博士在 综合分子结构教育和研究中心（IMSERC）的一套 NMR光谱技术，和Kelleher组质谱将促进携带 出这项研究。这些培训机会与合作和科学相结合， 西北大学的环境将确保拟议研究的成功， 将提供一个独立的，跨学科的研究生涯良好的准备。

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.