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由一个高度保守的 核糖核蛋白酶称为核糖体。核糖体的原子分辨率晶体结构， 4年前的报道，对我们理解蛋白质合成产生了革命性的影响， 但仍有许多有趣的问题可以通过晶体学来解决。研究 未来五年的计划有两个组成部分。首先，我们打算利用晶体学和遗传学 已经开发的工具，用于Haloarcula marismortui（Hma）获得核糖体的新见解 结构和功能。第二，核糖体和核糖体亚基的晶体将从新的 这样，关于核糖体结构和功能的问题就可以得到解答，而这些问题是无法解决的。 使用任何现有的核糖体晶体。 具体地说，假设众所周知的肽基转移酶性质的差异 来自不同物种的核糖体的中心是由其核苷酸之间的相互作用引起的。 将在Hma大核糖体中测试具有更远的非保守核苷酸的保守核心 亚基的遗传学和结晶学的结合。同样的工具也将用于确定 肽基转移酶中心如何在构象上响应高度保守碱基的突变， 肽基转移酶中心。此外，还将进行一系列实验， 制备在其出口通道中具有新生肽大亚基，并解析其结构 晶体学上。最终目标是制备真核核糖体晶体， 原子分辨率和它们的结构的确定。