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

使用位点特异性标记蛋白质进行核糖核酸酶 A 的时间分辨 NMR 折叠研究

• 批准号: 9634636
• 负责人: Deborah Wuttke
• 金额: $ 4万
• 依托单位: University of Colorado at Boulder
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-06-01 至 1998-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9634636&HistoricalAwards=false
• 关键词: Time; Resolved; NMR; Folding; Studies

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.

这是给位于博尔德的科罗拉多大学的启动资金 支持Deborah S.伍德克 研究 聚焦于非结构化多肽折叠成 蛋白质结构 时间分辨核磁共振技术 共振光谱学被用于研究在真实的时间的途径 核糖核酸酶A的折叠和展开。 通过特定地点， 核糖核酸酶A的N-末端区域中的同位素标记的残基， 这项研究利用了一维核磁共振的能力， 光谱学提供蛋白质中特定位点的信息， 折叠的过程和规避转让的困难， 分辨率 每种物质都被很好地分辨出来， 和交叉扬声器线形，作为当地结构环境的指标 和动力学性质，在整个动力学过程中， 过程 这项研究对于从根本上理解蛋白质具有重要意义 折叠和核糖核酸酶A的形成机制，蛋白质 催化单链核糖核酸的切割， 维持生命的遗传过程。