Structure and function of metabolic enzyme assemblies

代谢酶组装体的结构和功能

• 批准号: 10621612
• 负责人: Justin M Kollman
• 金额: $ 30.83万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-05-01 至 2028-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10621612
• 关键词: Amino Acids; Biochemical; Biological; Biophysics; Cells; Characteristics; Coupled; Cryoelectron Microscopy; Data; Disease; Energy Metabolism; Environment; Enzymes; Filament; In Vitro; Individual; Life; Medical; Metabolic; Metabolic Control; Metabolism; Molecular; Nucleotide Biosynthesis; Nutrient; Pathway interactions; Play; Polymers; Post-Translational Protein Processing; Process; Publishing; RNA Splicing; Reaction; Regulation; Role; Structure; System; Techniques; Variant; Work; enzyme activity; genetic regulatory protein; insight; polymerization; reconstitution; response; success

PROJECT SUMMARY Intermediate metabolism must be finely tuned, carefully balanced, and robustly adaptable to changes in envi- ronmental conditions. While the individual enzymes that drive most metabolic processes are well understood, only have we appreciated the widespread role of metabolic enzyme assemblies in metabolic organization and control. In particular, two types of structures, metabolic filaments that assemble from single enzyme types, and metabolons that co-assemble multiple enzymes in single pathways, provide important mechanisms for regulating enzyme activity and metabolic flux. Both filaments and metabolons assemble dynamically in cells, and alter the functions of the constituent enzymes to adapt to metabolic demand. This proposal builds on recent successes from our group describing the molecular mechanisms and functional consequences of enzyme assembly in mul- tiple systems, including nucleotide biosynthesis, energy metabolism, and amino acid regulation. Recent work, including our own published and preliminary data, shows that enzyme assembly is a general mechanism of control, and that assembly itself is controlled by cells in multiple ways, including by the levels of specific metab- olites, by posttranslational modification, by expression of splice variants that alter polymerization characteristics, and by interaction with regulatory proteins. We use in vitro reconstitution and cryo-electron microscopy to deter- mine the structural basis for assembly and regulation, coupled with biochemical, biophysical, and cell biological techniques in an integrative approach to understanding the functions of metabolic assemblies. This work will provide insight into the specific roles assembly plays in modulating the function of multiple enzymes, and illumi- nate general principles of metabolic control.

项目总结 中间代谢必须微调，仔细平衡，并有力地适应环境的变化-- 精神状况。虽然驱动大多数新陈代谢过程的单个酶已经很好地理解了， 只有我们认识到代谢酶组件在代谢组织和代谢过程中的广泛作用 控制力。具体地说，两种类型的结构，由单一酶类型组装的代谢细丝，以及 在单一途径中共同组装多种酶的代谢物提供了重要的调节机制 酶活性和代谢流量。细丝和代谢物都在细胞内动态组装，并改变 组成酶的功能，以适应代谢需求。这项建议是在最近的成功基础上提出的。 从我们的小组描述的分子机制和功能后果的酶组装在多- 三重系统，包括核苷酸生物合成、能量代谢和氨基酸调节。最近的工作， 包括我们自己发表的和初步的数据，表明酶组装是一种普遍的机制 控制，而该程序集本身由细胞以多种方式控制，包括由特定代谢抗体的水平控制。 通过翻译后修饰，通过表达改变聚合特性的剪接变体， 并通过与调节蛋白的相互作用。我们使用体外重建和冷冻电子显微镜来阻止- 挖掘组装和调节的结构基础，再加上生化、生物物理和细胞生物学 以综合的方法了解代谢组合的功能的技术。这项工作将 深入了解组件在调节多种酶的功能中所起的具体作用，以及 新陈代谢控制的一般原则。