Time-Resolved NMR Folding Studies of Ribonuclease A Using Site-Specifically Labeled Protein

这项授予科罗拉多大学博尔德分校的启动奖学金支持了黛博拉·S·伍特克教授的研究。这项研究的重点是非结构多肽折叠成蛋白质结构的机制。利用时间分辨核磁共振波谱技术实时研究核糖核酸酶A的折叠和去折叠途径，通过定位于核糖核酸酶A N末端的同位素标记残基，利用一维核磁共振波谱在折叠过程中提供蛋白质特定位置的信息的能力，避免了分配和分辨的困难。每个物种都被很好地分辨，允许化学位移和相声线形作为局部结构环境和动力学性质的指示器，在整个动力学过程中被跟踪。这项研究对于基本了解蛋白质折叠和核糖核酸酶A的形成机制具有重要意义。核糖核酸酶A是一种在维持生命的遗传过程中催化单链核糖核酸切割的蛋白质。

This starter grant award to the University of Colorado at Boulder supports the research of Professor Deborah S. Wuttke. The research focusses on the mechanism by which unstructured polypeptides fold into protein structures. The technique of time-resolved nuclear magnetic resonance spectroscopy is employed for study in real time of the pathways of folding and unfolding in ribonuclease A. Through site-specifically, isotopically labeled residues in the N-terminal region of ribonuclease A, the study takes advantage of the ability of 1D nuclear magnetic resonance spectroscopy to provide information on specific sites in a protein during the course of folding and circumvents the difficulties of assignment and resolution. Each species is well-resolved, allowing the chemical shift and crosspeak lineshapes, as indicators of local structural environment and dynamical properties, to be followed throughout the entire kinetic process. The research is important in the fundamental understanding of protein folding and the mechanism of formation of ribonuclease A, the protein that catalyzes the cleavage of single-stranded ribonucleic acid in the course of life-sustaining genetic processes.