Structural Basis of Metallocofactor Delivery and Repair

金属辅因子传递和修复的结构基础

• 批准号: 10054102
• 负责人: Francesca Vaccaro
• 金额: $ 4.6万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10054102
• 关键词: Binding; Biological Models; Chemistry; Chimeric Proteins; Cobalamin; Coenzymes; Complex; Crystallization; Data; Disease; Electron Microscopy; Electron Spin Resonance Spectroscopy; Electrons; Enzymes; Excision; GTP-Binding Protein alpha Subunits, Gs; GTP-Binding Proteins; Guanosine Triphosphate; Guanosine Triphosphate Phosphohydrolases; Health; Human; Hydrolysis; Laboratories; Lead; Length; Mediating; Metabolism; Methylmalonyl-CoA Mutase; Methylobacterium extorquens; Molecular; Molecular Chaperones; Molecular Conformation; Molecular Mechanisms of Action; Molecular Sieve Chromatography; Mutase; Mutation; Negative Staining; Nucleotides; Pathway interactions; Play; Process; Proteins; Roentgen Rays; Role; Sampling; Spin Labels; Structure; Substrate Domain; System; Tertiary Protein Structure; Three-Dimensional Image; Time; Vitamin B 12; Work; X-Ray Crystallography; analog; biophysical techniques; cobamamide; cofactor; human disease; image reconstruction; insight; isobutyryl-coenzyme A; methylmalonic aciduria; polypeptide; protein activation; protein complex; repaired

PROJECT SUMMARY/ABSTRACT Complex and reactive metallocofactors are often responsible for challenging chemistry that is critical to cellular metabolism. It is therefore not surprising that the transport of these cofactors to their target enzymes often involves multiple proteins and complex delivery pathways. Mutations in any of these components may result in a breakdown of the pathway causing deficits in the overall activity of the target enzyme and leading to a disease state, as in the case with the vitamin B12 (cobalamin, Cbl) delivery pathway. In this proposal, we will study enzymes from humans, Methylobacterium extorquens and Cupriavidus metallidurans in order to determine the molecular mechanisms of Cbl transport and installation. Our approach involves the use of a variety of biophysical methods that will allow us to visualize the interactions between a G-protein chaperone and its target Cbl- dependent mutase in the final step of the Cbl delivery pathway. This work will inform on the structural basis of chaperone-mediated delivery and repair of Cbl. By combining X-ray crystallography, electron microscopy and electron paramagnetic resonance spectroscopy, we will explore the molecular mechanism of action of the G- protein chaperones involved in the installation and repair of adenosylcobalamin in their target mutases.

项目摘要/摘要 复杂和反应性的金属生物活性剂通常负责挑战化学，这对细胞至关重要 代谢。因此，毫不奇怪，这些辅助因子经常向其靶酶运输 涉及多种蛋白质和复杂的递送途径。这些组件中的任何一个中的突变都可能导致 途径的分解导致靶酶的整体活性缺陷并导致疾病 与维生素B12（钴胺，CBL）递送途径一样，状态。在此提案中，我们将学习 来自人类的酶，八甲基甲虫和杯状金属尿素，以确定 CBL传输和安装的分子机制。我们的方法涉及使用各种生物物理 将使我们能够可视化G蛋白伴侣伴侣及其靶标CBL之间的相互作用的方法 在CBL递送途径的最后一步中，依赖性突变酶。这项工作将以结构性为基础 伴侣介导的CBL的递送和修复。通过结合X射线晶体学，电子显微镜和 电子顺磁共振光谱学，我们将探索G-的分子作用机理 蛋白质伴侣参与其靶突变中腺苷白素的安装和修复。