Characterization of new methanobactins and their biosynthetic enzymes

项目摘要/摘要 甲烷结合蛋白(MBN)是由核糖体合成和翻译后修饰的 甲烷氧化细菌产生的多肽天然产物的获得 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)。以前描述的MBN 含有具有两个杂环/硫代酰胺对的多肽主链，提供两个氮和 铜(I)的两个硫配体。这些MBN结合铜具有高亲和力和特异性，并 正在研究中作为治疗人类铜代谢疾病的候选药物。近期 生物信息学研究表明，有一些新类型的MBN被预测为 在结构和化学性质上不同于先前表征的MBN。此外， 几种未确定特征的酶，有望参与这些酶的生物合成 新的MBN还没有被很好地理解。在这个项目中，未刻画的MBN与预测的小说 将探索其结构，并将研究负责其生物合成的酶 体内和体外方法都是如此。这项研究将为我们提供对 MBN结构和功能的多样性，将阐明新的酶化学，并将 影响MBN和MBN类分子作为治疗药物的发展。 已经计划了一个多方面的培训计划。各种生化方面的培训 以及分离和分离所需的遗传和光谱/光谱技术 罗森茨韦格实验室将提供对MBN及其生物合成酶的表征。 此外，在实验室以外的正规培训机会，如在生活计划 高级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 这两个研讨会都与拟议的研究密切相关，将会举行。这个 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 电子顺磁共振(EPR)谱霍夫曼实验室，张博士在 综合分子结构教育与研究中心(IMSERC) 核磁共振波谱技术，以及用于质谱学的凯莱赫基团，将便于携带 在这项研究之外。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的研究和 将为独立的、跨学科的研究生涯提供极好的准备。

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.