Protein-Protein Interactions in Natural Product Biosynthesis

天然产物生物合成中的蛋白质-蛋白质相互作用

• 批准号: 10249686
• 负责人: Michael D. Burkart
• 金额: $ 1.29万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-03-01 至 2024-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10249686
• 关键词: Anabolism; Antibiotics; Antifungal Agents; Artificial Arm; Biology; Carrier Proteins; Chemicals; Development; Environment; Escherichia coli; Fatty-acid synthase; Feedback; Fluorescent Dyes; Goals; Isotope Labeling; Knowledge; Length; Ligands; Location; Medicine; Natural Products; Nuclear Magnetic Resonance; Pathway interactions; Production; Proteins; Route; Site; Structure; System; Tertiary Protein Structure; Therapeutic; Time; anti-cancer; enoyl reductase; functional group; novel; peptide synthase; polyketide synthase; protein protein interaction; tool

Project Summary This project aims to develop a new synthetic tool to investigate the ligand·protein and protein·protein interactions in carrier protein-dependent biosynthetic pathways including fatty acid synthase (FAS), polyketide synthase (PKS), and non-ribosomal peptide synthetase pathways (NRPS). The natural products produced from these pathways constitute a majority of therapeutics ranging from antibiotics and antifungals to anticancer compounds. Manipulating and controlling the biosynthesis of these pathways toward the production of new bioactive compounds has been a long-term goal of the field. Developing an understanding of the protein·protein interactions will allow us to make strides toward these long-term goals, as interactions between the carrier protein and partner protein domains guide the processivity and reactivity of the pathway. In the Burkart lab, we have previously developed chemical biology tools to study snapshots of these interactions or visualize them through the use of fluorescent dyes. It is proposed in this project to create a new site-specific 15N isotopically labeled tool that will be able to visualize changes in the environment of the ligand attached to the carrier protein and give information about chain flipping of this prosthetic arm through 15N nuclear magnetic resonance (NMR) studies. This tool will be able to mimic the native prosthetic arm while giving real time feedback on the temporal location of the probe and its structural surroundings in a simplified form. This versatile tool will be able to give information on ligand·protein interactions and chain flipping phenomena in multiple systems through its adaptable synthetic route including Escherichia coli AcpP with the chain-length specific LipB, and with the functional group specific ketoreductase and enoyl reductase E. coli FAS domains.

项目概要 该项目旨在开发一种新的合成工具来研究配体·蛋白质和 载体蛋白依赖性生物合成途径（包括脂肪酸）中蛋白质·蛋白质相互作用 合酶 (FAS)、聚酮化合物合酶 (PKS) 和非核糖体肽合成酶途径 （国家公共服务计划）。这些途径产生的天然产物构成了大部分治疗药物 范围从抗生素和抗真菌剂到抗癌化合物。操纵和控制 这些生产新生物活性化合物的途径的生物合成已被 该领域的长期目标。加深对蛋白质·蛋白质相互作用的理解将 由于载体蛋白之间的相互作用，使我们能够朝着这些长期目标迈进 和伙伴蛋白结构域指导该途径的持续性和反应性。在布卡特 实验室，我们之前开发了化学生物学工具来研究这些相互作用的快照 或通过使用荧光染料将它们可视化。建议在该项目中创建一个新的 特定位点的 15N 同位素标记工具将能够可视化环境变化 附在载体蛋白上的配体的信息，并提供有关该配体链翻转的信息 假臂通过 15N 核磁共振 (NMR) 研究。这个工具将能够 模仿原生假肢，同时提供假肢时间位置的实时反馈 探头及其周围结构的简化形式。这个多功能工具将能够提供 多系统中配体·蛋白质相互作用和链翻转现象的信息 通过其适应性强的合成路线，包括具有链长特异性的大肠杆菌 AcpP LipB，并具有功能组特异性酮还原酶和烯酰还原酶大肠杆菌 FAS 域。