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: 10548747
• 负责人: Michael D. Burkart
• 金额: $ 46.88万
• 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 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10548747
• 关键词: Acyl Carrier Protein; Acyltransferase; Address; Anabolism; Antibiotics; Antifungal Agents; Antineoplastic Agents; Biological; Biological Assay; Biology; Book Chapters; Carrier Proteins; Chemicals; Communication; Computer Models; Computing Methodologies; Crosslinker; Data; Development; Engineering; Enzymes; Escherichia coli; Experimental Designs; Fatty Acids; Fatty-acid synthase; Foundations; Funding; Goals; Hybrids; In Vitro; Informatics; Kinetics; Knowledge; Laboratories; Length; Libraries; Macromolecular Complexes; Maps; Medicine; Metabolic; Metabolic Pathway; Molecular; Multi-Drug Resistance; Mutagenesis; Mutation; NMR Spectroscopy; Natural Products; Nature; Oxidases; Oxidoreductase; Pathway interactions; Peptides; Phase; Physical condensation; Play; Post-Translational Protein Processing; Production; Productivity; Progress Reports; Property; Protein Engineering; Protein-Protein Interaction Map; Proteins; Publications; Publishing; Refractory; Research; Resolution; Role; Specificity; Structure; System; Tertiary Protein Structure; Therapeutic; Thermodynamics; Work; X-Ray Crystallography; analog; bacterial resistance; clay; crosslink; design; drug development; drug discovery; engineering design; enoyl reductase; fatty acid biosynthesis; human disease; improved; in silico; in vivo; information gathering; innovation; interest; microbial; molecular dynamics; natural product hybrids; next generation; novel therapeutics; peptide synthase; polyketide synthase; polyketides; programs; protein complex; protein crosslink; protein protein interaction; secondary metabolite; structural biology; synthetic biology; tool; tool development

7. Project Summary. This program investigates the role of protein'protein interactions in carrier protein-dependent biosynthesis, including fatty acid synthase, polyketide synthase, and non-ribosomal peptide synthetase pathways. The natural products associated with these pathways serve as therapeutics including antibiotics, antifungals, and anticancer agents. The ability to direct the biosynthesis of these pathways for the production of untried bioactive compounds is of critical importance for new and improved therapies. In prior years, we demonstrated how protein'protein interactions between carrier proteins and partner protein domains direct reactivity and guide processivity. Moreover, we developed a suite of chemical biology tools to stabilize and interrogate these interactions at atomic resolution through innovative crosslinker development and structural biology. We now propose to expand these tools with new, caged crosslinkers designed to capture transient partner proteins. With this comprehensive library of crosslinkers, we will evaluate carrier protein interactions with partner proteins that include ketoreductases, enoyl reductases, and thioesterases from fatty acid and polyketide synthases, ketosynthase/chain-length factors and acyltransferases from polyketide synthases, and condensation domains, halogenases, and oxidases from non-ribosomal peptide synthetases. Once captured, these crosslinked species will be studied by solution-phase NMR, kinetic and thermodynamic assays, X-ray crystallography, and molecular dynamics simulations to more fully elucidate mechanism and specificity conferred by carrier protein-substrate/intermediate/produ interactions. Finally, we will use data collected in this program as an informatic platform to enable synthetic biological production of new natural product hybrids.

7。项目摘要。 该程序研究了蛋白质蛋白相互作用在载体蛋白依赖性生物合成中的作用， 包括脂肪酸合酶，聚酮化合物合酶和非核糖体肽合成酶途径。这 与这些途径相关的天然产物作为治疗药，包括抗生素，抗真菌剂和 抗癌剂。将这些途径的生物合成指导未经尝试的生物合成的能力 生物活性化合物对于新的和改进的疗法至关重要。在前几年，我们证明了 载体蛋白与伴侣蛋白结构域之间的蛋白质蛋白质相互作用如何直接反应性和 指导过程。此外，我们开发了一套化学生物学工具来稳定和询问这些工具 通过创新的交联剂开发和结构生物学，在原子分辨率下进行的相互作用。我们现在 建议使用旨在捕获瞬态合作伙伴蛋白的新的，笼中的交联器扩展这些工具。 使用这个全面的交联库，我们将评估载体蛋白与合作伙伴的相互作用 包括酮酸和脂肪酸和聚酮化合物的蛋白质，包括酮糖酶，烯酰还原酶和硫酯酶 合成酶，酮合酶/链长因子和来自聚酮化合物合酶的酰基转移酶，以及 来自非核糖体肽合酶的冷凝域，卤代酶和氧化酶。一旦被捕获， 这些交联物种将通过溶液相NMR，动力学和热力学测定，X射线研究 晶体学和分子动力学模拟，以更完全阐明机制和特异性 由载体蛋白质 - 基底/中间/产品相互作用赋予。最后，我们将使用其中收集的数据 作为一个信息平台的程序，可以使新天然产品混合动力的合成生物生产。