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

基本信息

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
项目类别：
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&apos -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

项目摘要

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.

项目概要微生物产生结构多样的分子，其中许多已被成功地重新利用人类作为药剂。这些分子是由多酶组装体制造的，使用酰基载体蛋白（ACP）来修饰和转移化学中间体。自然的合理再设计酶组装体为生产新抗生素提供了一条令人兴奋的可能途径，但任何方法的成功重新设计方法取决于对导致化学生产混合 ACP- 的原因的透彻理解酶的相互作用。本研究的目的是了解 ACP 如何与其分子相互作用货物和伙伴酶。 ACP 的动态结构及其蛋白质的瞬时性质相互作用使得这类蛋白质的研究特别具有挑战性。因此，ACP 相互作用将是使用特定位点振动光谱和沉降速度实验观察到，一种创新的对快速构象变化和弱蛋白质-蛋白质相互作用敏感的方法。信息这项工作将在从传统方法获得的数据的背景下进行分析，以确定能够产生新颖结构分子的候选杂合 ACP-酮合酶伴侣。的能力将评估生产聚酮化合物的杂化对。这些研究的结果将指导未来具有潜在药物活性的新型小分子的生物合成。