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: 10540355
• 负责人: Francesca Vaccaro
• 金额: $ 2.96万
• 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 2023-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10540355
• 关键词: Binding; Biological Models; Chemistry; Chimeric Proteins; Cobalamin; Coenzymes; Complex; Data; Disease; Electron Microscopy; Electron Spin Resonance Spectroscopy; Electrons; Enzymes; Excision; 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; Process; Proteins; Roentgen Rays; Role; Sampling; Spin Labels; Structure; Substrate Domain; System; Tertiary Protein Structure; Three-Dimensional Image; Time; Visualization; 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; 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 复杂的和反应性的金属相关因子常常对对细胞至关重要的化学构成挑战。 新陈代谢。因此，不足为奇的是，这些辅因子运输到他们的目标酶经常 涉及多个蛋白质和复杂的传递途径。这些成分中的任何一个的突变都可能导致 导致目标酶的整体活性不足并导致疾病的途径的崩溃 状态，与维生素B12(钴胺，Cbl)传递途径的情况一样。在这项建议中，我们将研究 从人、甲烷杆菌和金属铜绿假单胞菌中提取的酶以确定 Cbl转运和安装的分子机制。我们的方法涉及到使用各种生物物理 这些方法将使我们能够可视化G蛋白伴侣与其靶Cbl之间的相互作用。 在Cbl传递途径的最后一步中依赖的变位酶。这项工作将在结构基础上提供信息 伴侣介导的Cbl的传递和修复。通过将X射线结晶学、电子显微镜和 电子顺磁共振波谱，我们将探索G-的分子作用机制。 蛋白质伴侣参与腺苷钴胺在其靶标变位酶中的安装和修复。