The Mechanism by Which Ribosomes Form Proteins

这项提议的目标是从分子力学的角度了解核糖体及其相互作用的大分子是如何形成蛋白质的。具体地说，这项拟议的研究是为了确定在核糖体上拉长多肽的反应步骤中，tRNAs和mRNAs如何相互移动以及与核糖体结构成分的关系。该方法是将荧光标记的tRNAs或mRNAs结合到标记或未标记的核糖体上，然后测量多肽延长周期完成特定反应前后的各种物理变化。探针被共价连接在tRNA、mRNA类似物、核糖体蛋白和核糖体RNA上的特定已知位置。后者被重新组装成具有功能的核糖体。所有的蛋白质都是在核糖体上与mRNA和tRNA一起形成的，其机制似乎与所有生物体相似。因此，对核糖体结构和功能的了解是理解蛋白质是如何在分子水平上形成的基础，蛋白质是催化生命系统中发生的无数反应的。

The goal of this proposal is to understand in molecular mechanical terms how ribosomes and the macromolecules that interact with them function to form proteins. Specifically, the proposed research is to determine how tRNAs and mRNA move in relation to each other and to ribosomal structural components during the reaction steps by which peptides are elongated on ribosomes. The appraoch is to bind fluorescently labeled tRNAs or mRNA to labeled or unlabeled ribosomes, then to measure various physical changes before and after completion of a specific reaction of the peptide elongation cycle. Probes are covalently attached at specific known sites on tRNA, mRNA analogues, ribosomal proteins and ribosomal RNA. The latter are reassembled into functionally competent ribosomes. All proteins are formed on ribosomes with mRNA and tRNA by a mechanism that appears to be similar for all organisms. Thus, knowledge of ribosome structure and function is fundamental to an understanding of how proteins, which catalyze the myriad reactions that occur in living systems, are formed at the molecular level.