Elucidating the Mechanism of the Ribosome

阐明核糖体的机制

• 批准号: 6946838
• 负责人: SHENG-LIN HUANG
• 金额: $ 4.83万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2006-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6946838
• 关键词: biochemistry; catalyst; chemical conjugate; chemical kinetics; chemical structure function; enzyme mechanism; enzyme substrate; genetic translation; immunoprecipitation; nucleic acids; peptides; postdoctoral investigator; protein biosynthesis; ribosomal RNA; ribosomes; ribozymes; small molecule; stereochemistry; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): This proposal outlines a series of bioorganic studies to characterize the mechanism of rRNA catalyzed protein synthesis. The project involves the preparation of novel peptide nucleic acid conjugates and transition state analogues that will be used to achieve the following goals: (i) Define the atomic interactions within the ribosome active site when A- and P- substrates are bound, but prior to the chemical step. (ii) Determine if the macroscopic pKa of the peptidyl transfer reaction corresponds to the pKa of the nucleophile under conditions where chemistry is the limiting step in the reaction. (iii) Determine the atomic interactions and the affinity of chiral inhibitors that mimic the tetrahedral intermediate. These inhibitors will illuminate the interactions between the ribozyme and the substrates that elicit the catalytic power required of the cellular peptide synthesis machinery. The study will provide a fundamental understanding of translation and the mechanisms of RNA catalysis; processes that are fundamentally important to cell growth and development.

描述(由申请人提供)： 这项建议概述了一系列生物有机研究，以表征rRNA催化蛋白质合成的机制。该项目包括制备新型多肽核酸结合物和过渡态类似物，用于实现以下目标：(I)确定当A-和P-底物结合时但在化学步骤之前核糖体活性部位内的原子相互作用。(2)确定在化学是反应的限制步骤的条件下，缩氨酸转移反应的宏观pKa是否对应于亲核试剂的pKa。(Iii)确定模拟四面体中间体的手性抑制剂的原子相互作用和亲和力。这些抑制剂将阐明核酶和底物之间的相互作用，从而产生细胞多肽合成机器所需的催化能力。这项研究将提供对翻译和RNA催化机制的基本理解；这些过程对细胞的生长和发育至关重要。