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.

项目总结