Capturing the key protein and substrate interactions in polyketide synthases using isosteric mimetics

使用等排模拟物捕获聚酮合酶中的关键蛋白质和底物相互作用

• 批准号: 10223911
• 负责人: Rebecca N. Re
• 金额: $ 3.89万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN DIEGO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10223911
• 关键词: 3-hydroxybutanal; Active Sites; Acyl Carrier Protein; Anabolism; Anti-Bacterial Agents; Antibiotic Resistance; Antibiotics; Antifungal Agents; Antineoplastic Agents; Attention; Biochemistry; Biological; Biology; Carbon; Carrier Proteins; Chemicals; Coenzyme A; Complex; Crystallization; Custom; Development; Disease; Doxorubicin; Engineering; Enzymes; Erythromycin; Goals; Health; Immunosuppressive Agents; Isoxazoles; Knowledge; Lead; Learning; Length; Malonyl Coenzyme A; Modeling; Modern Medicine; Molecular; Molecular Machines; Natural Products; Nature; Organism; Paper; Pharmaceutical Preparations; Physical condensation; Process; Production; Proteins; Publishing; Reaction; Research; Research Personnel; Side; Sirolimus; Specificity; Structure; Structure-Activity Relationship; Substrate Interaction; Synthesis Chemistry; System; Therapeutic; Training; Vertebral column; Work; X-Ray Crystallography; actinorhodin; analog; clay; crosslink; design; diphenyl; enzyme activity; enzyme pathway; enzyme structure; falls; interdisciplinary approach; mimetics; novel; novel anticancer drug; novel therapeutics; pharmacophore; polyketide synthase; prevent; protein complex; protein crosslink; protein protein interaction; structural biology; success; symposium; thioether; tool

Project Summary. With the majority of therapeutic drugs available on the market being natural products or derivatives of them, understanding how organisms and enzymes function to produce these structurally complex compounds is essential. Polyketides are a class of secondary metabolites that are biosynthesized by polyketide synthases (PKSs) and often serve as antibacterial, antifungal, and anticancer agents. The PKSs are complex biological machineries that involve proteins and substrates interacting with one another with high specificity to assemble polyketides. These unique protein-protein and protein-substrate interactions are the basis for how these synthases are governed and are therefore critical to understand. Common in all three types of PKSs is the iterative elongation of polyketide intermediates by two-carbon units, but how their respective elongation enzymes function and stabilize the substrates while preventing them from undergoing unwanted side reactions continues to remain unknown. In this proposal, we aim to first understand the fit of growing polyketones in the pocket of a carrier protein-guided elongation enzyme by developing isosteric mimetics of polyketide intermediates from a type II PKS model. Here, we will use crosslinking to trap the partner proteins to elucidate the key interactions as the intermediates are elongated. We then plan to apply similar chemical biology tools in our second research aim to define the substrate interactions catalyzed by a CoA-dependent elongation enzyme in a type III PKS system. In this study, we will develop polyketide intermediate mimetics and malonyl-CoA analogs to be able to provide a snapshot of the natural substrate interactions through x-ray crystallography. These studies will allow us to uncover the molecular details that drive the elongation process responsible for building the core carbon backbone of polyketides. Gaining a deeper understanding of these protein and substrate interactions enables their manipulation and redesign to produce novel polyketides with different pharmacophores.

项目摘要。市场上的大多数治疗药物都是天然产物或 它们的衍生物，了解生物体和酶如何发挥作用来产生这些结构复杂的 化合物是必不可少的。聚酮化合物是一类由聚酮化合物生物合成的次级代谢产物 合成酶 (PKS)，通常用作抗菌剂、抗真菌剂和抗癌剂。 PKS 很复杂 涉及蛋白质和底物以高度特异性相互作用的生物机器 组装聚酮化合物。这些独特的蛋白质-蛋白质和蛋白质-底物相互作用是如何 这些合酶受到控制，因此理解它们至关重要。所有三种类型的 PKS 的共同点是 聚酮化合物中间体通过两个碳单元的迭代延伸，但它们各自的延伸酶如何 发挥作用并稳定底物，同时防止它们发生不需要的副反应 保持未知。在本提案中，我们的目标是首先了解在口袋里种植聚酮的适合性 通过开发来自聚酮化合物中间体的等排模拟物，载体蛋白引导的延伸酶 II 型 PKS 模型。在这里，我们将使用交联来捕获伴侣蛋白，以阐明关键的相互作用： 中间体被拉长。然后我们计划在我们的第二项研究中应用类似的化学生物学工具 旨在定义 III 型 PKS 中 CoA 依赖性延伸酶催化的底物相互作用 系统。在这项研究中，我们将开发聚酮化合物中间模拟物和丙二酰辅酶A类似物，以便能够 通过 X 射线晶体学提供天然基质相互作用的快照。这些研究将允许 我们将揭示驱动构建核心碳的延伸过程的分子细节 聚酮化合物的骨架。更深入地了解这些蛋白质和底物相互作用使得 对它们进行操作和重新设计以生产具有不同药效团的新型聚酮化合物。