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.

项目摘要 中间代谢必须精细调整，仔细平衡，并有力地适应环境的变化。 环境条件虽然驱动大多数代谢过程的单个酶已经被很好地理解， 只有我们意识到代谢酶组装在代谢组织中的广泛作用， 控制特别是，两种类型的结构，从单一酶类型组装的代谢丝， 在单一途径中共同组装多种酶的代谢子，为调节 酶活性和代谢通量。纤维和代谢子在细胞中动态组装，并改变细胞的结构。 组成酶的功能，以适应代谢的需求。这一建议是在最近成功的基础上提出的 来自我们的小组，描述了穆尔中酶组装的分子机制和功能后果， 三个系统，包括核苷酸生物合成、能量代谢和氨基酸调节。近期工作、 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，而组装本身是由细胞以多种方式控制的，包括特定的代谢水平， 通过翻译后修饰，通过表达改变聚合特征的剪接变体， 以及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确定- 挖掘组装和调节的结构基础，再加上生物化学，生物物理学和细胞生物学 技术在一个综合的方法来理解代谢组件的功能。这项工作将 提供深入了解组装在调节多种酶的功能中所起的特定作用，并阐明 代谢控制的基本原则。