Functional Hybrid Natural Product Synthases by Tracking Acyl Carrier Protein Binding and Conformational Dynamics

通过跟踪酰基载体蛋白结合和构象动力学进行功能性杂化天然产物合成

• 批准号: 9171419
• 负责人: Louise Karine Charkoudian
• 金额: $ 39.03万
• 依托单位: HAVERFORD COLLEGE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9171419
• 关键词: 4' -phosphopantetheine; Active Sites; Acyl Carrier Protein; Affect; Anabolism; Antibiotics; Automobile Driving; Binding; Binding Proteins; Catalysis; Chemicals; Chemistry; Complement; Complex; Data; Environment; Enzyme Interaction; Enzymes; Event; Family; Foundations; Future; Goals; Hybrids; Knowledge; Label; Length; Methodology; Methods; Molecular; Natural Products; Nature; Nitriles; Outcome; Pathway interactions; Pharmacologic Substance; Positioning Attribute; Process; Production; Productivity; Proteins; Reporting; Role; Route; Sampling; Site; Spectrum Analysis; Stretching; Structure; Sulfhydryl Compounds; Testing; Thiocyanates; Variant; Work; arm; base; design; high throughput screening; innovation; insight; intermolecular interaction; microorganism; novel; protein function; protein protein interaction; protein structure; protein structure function; research study; sedimentation velocity; small molecule; success; temporal measurement; tool

PROJECT SUMMARY Microorganisms produce structurally diverse molecules, many of which have been successfully repurposed by mankind as pharmaceutical agents. These molecules are manufactured by multi-enzyme assemblies, which use acyl carrier proteins (ACPs) to modify and transfer chemical intermediates. Rational redesign of natural enzyme assemblies presents an exciting possible route to produce new antibiotics, but the success of any redesign approach depends on a thorough understanding of what leads to chemically productive hybrid ACP- enzyme interactions. The goal of this study is to understand how ACPs interact with both their molecular cargoes and partner enzymes. The dynamic structures of ACPs and the transient nature of their protein interactions make this class of proteins particularly challenging to study. Therefore, ACP interactions will be observed using site-specific vibrational spectroscopy and sedimentation velocity experiments, an innovative approach that is sensitive to fast conformational changes and weak protein-protein interactions. Information from this work will be analyzed in the context of data acquired from traditional methodologies to identify candidate hybrid ACP-ketosynthase partners capable of producing molecules of novel structure. The ability of the hybrid pairs to produce polyketides will be evaluated. Results from these studies will guide the future biosynthesis of novel small molecules with potential pharmaceutical activity.

项目总结