Program in Macromolecular Structure, Motion, Control

高分子结构、运动、控制程序

• 批准号: 7297743
• 负责人: PETER B. MOORE
• 金额: $ 18.02万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-25 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7297743
• 关键词: Halobacteriaceae; X ray crystallography; aminoacyltransferase; chemical structure; conformation; eukaryote; intermolecular interaction; macromolecule; molecular dynamics; ribosomes; species difference

Messenger RNA-directed protein synthesis is catalyzed in all cells by a highly conserved ribonucleoprotein enzyme called the ribosome. Atomic resolution crystal structures of ribosomes, the first of which were reported 4 years ago, have had a revolutionary impact on our understanding of protein synthesis, but many interesting questions remain that can best be approached crystallographically. The research proposed for the next 5 years has two components. First, we intend to use the crystallographic and genetic tools already developed here for Haloarcula marismortui (Hma) to obtain new insights into ribosome structure and function. Second, crystals of ribosomes and ribosomal subunits will be prepared from new species so that questions about ribosome structure and function can be answered that cannot be addressed using any of the ribosome crystals now available. Specifically, the hypothesis that the well known differences in the properties of the peptidyl transferase centers of the ribosomes from different species are caused by interactions between the nucleotides in its conserved core with more remote, non-conserved nucleotides will be tested in the Hma large ribosomal subunit using a combination of genetics and crystallography. The same tools will also be used to determine how the peptidyl transferase center responds conformationally to the mutation of highly conserved bases in the peptidyl transferase center. In addition, a series of experiments will be carried out the objective of which is to prepare large subunit that have nascent peptides in their exit tunnels, and solve their structures crystallographically. The final goal is the preparation of crystals of eukaryotic ribosomes that diffract to atomic resolution, and the determination of their structures.

信使RNA指导的蛋白质合成在所有细胞中均由高度保守的催化 核糖核蛋白酶称为核糖体。核糖体的原子分辨率晶体结构，第一个 四年前报道的，对我们对蛋白质合成的理解产生了革命性的影响， 但仍然存在许多有趣的问题，最好从晶体学的角度来解决这些问题。研究 未来 5 年的提议有两个组成部分。首先，我们打算利用晶体学和遗传学 这里已经为 Haloarcula marismortui (Hma) 开发了工具，以获得对核糖体的新见解 结构和功能。其次，核糖体和核糖体亚基的晶体将从新的 物种，以便能够回答有关核糖体结构和功能的无法解决的问题 使用现有的任何核糖体晶体。 具体来说，假设肽基转移酶的特性存在众所周知的差异 不同物种的核糖体中心是由其核苷酸之间的相互作用引起的 具有更远程、非保守核苷酸的保守核心将在 Hma 大核糖体中进行测试 亚基结合遗传学和晶体学。相同的工具也将用于确定 肽基转移酶中心如何对高度保守碱基的突变做出构象反应 肽基转移酶中心。此外，还将进行一系列实验，其目的是 是制备在其出口通道中具有新生肽的大亚基，并解析其结构 晶体学上。最终目标是制备可衍射的真核核糖体晶体 原子分辨率及其结构的测定。