Conformational changes of the multi-modular MetH

多模块 MetH 的构象变化

• 批准号: 7493290
• 负责人: ALLISON C LAMANNA
• 金额: $ 2.52万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-09-01 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7493290
• 关键词: Binding; Catalysis; Chemistry; Cobalamin; Complex; Dancing; Data; Distant; Enzyme Kinetics; Enzymes; Equilibrium; Escherichia coli; Fellowship; Fluorescence; Fluorescence Resonance Energy Transfer; Folate; Homocysteine; Homocystine; Homologous Gene; Individual; Kinetics; Label; Ligands; Measures; Methionine; Molecular; Molecular Conformation; Monitor; Movement; Mutation; Names; Pathway interactions; Pliability; Protein Binding Domain; Protein Fragment; Proteins; Rate; Reaction; Reporting; Research; S-Adenosylmethionine; Spectrum Analysis; Structure; Study models; System; Tertiary Protein Structure; Thermotoga maritima; Vitamin B 12; X-Ray Crystallography; chemical kinetics; chemical reaction; cofactor; fluorophore; insight; methyl group; protein function; size

Many large, multi-domain proteins must undergo dramatic structural movements to perform their functions. X-ray crystallography provides static pictures of the protein but does not describe its movement. To more fully understand the function of these proteins, it is useful to study the conformational changes undergone by the protein and the rates of those changes. Fluorescence resonance energy transfer between fluorophore- labeled domains reports on the distances and distance change between those domains. The conformational changes of the enzyme cobalamin-dependent methionine synthase (MetH) can be studyied by labeling its 5 domains with different fluorophores to measure the interactions between the different domains. MetH is important in the pathway for the utilization of folate derivatives and homocysteine. During catalysis, MetH must undergo a complex molecular "dance" to allow each domain to interact with vitamin B12. MetH is an ideal candidate for the study of conformational changes because rates of conformational change can be compared to the previously reported kinetics, and it is known that the chemistry of the reaction is not rate- limiting. This research will provide insight for the study of additional multi-domain proteins and enzymes.

许多大型多域蛋白必须进行戏剧性的结构运动才能执行其功能。 X射线晶体学提供了蛋白质的静态图片，但没有描述其运动。更多 完全了解这些蛋白质的功能，研究构象变化的发生很有用 蛋白质和这些变化的速率。荧光荧光共振能量转移荧光团 标记的域报告了这些域之间的距离和距离变化。构象 可以通过标记其5来研究酶钴胺素依赖性蛋氨酸合酶（MET）的变化 具有不同荧光团的结构域测量不同域之间的相互作用。甲基是 在利用叶酸衍生物和同型半胱氨酸的途径中很重要。在催化过程中，甲基苯丙胺 必须进行复杂的分子“舞蹈”，以使每个结构域与维生素B12相互作用。甲基是一个 研究构象变化的理想候选者，因为构象变化的速率可以是 与先前报道的动力学相比，众所周知，反应的化学不合理 限制。这项研究将为研究其他多域蛋白和酶的研究提供见识。