T-4 Lysozyme as a Model for Protein Folding

T-4 溶菌酶作为蛋白质折叠的模型

• 批准号: 9206057
• 负责人: Frederick Dahlquist
• 金额: $ 35.4万
• 依托单位: University of Oregon Eugene
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 1996-01-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9206057&HistoricalAwards=false
• 关键词: Lysozyme; Model; Protein; Folding

The purpose of this proposal is to better understand the interrelationships of protein structure, stability, and dynamics using T4 lysozyme as a model. This moderate sized protein of 164 residues has a known 3-dimensional crystal structure. The protein has two structural domains and no natural disulfide bonds. Many mutants exist that modulate its thermodynamic stability and more than 100 mutant crystal structures have been determined by Brian Matthews' laboratory at the University of Oregon. We have assigned the amide resonances of the wild type and several mutant proteins using modern nuclear magnetic resonance techniques (NMR). We propose to use these methods to investigate (1) the ionization behavior of the 19 carboxyl groups of the protein with the goal of understanding electrostatic and polarity effects in the protein; (2) the dynamics of the folded state; (3) determine the role in the folding pathway of an early folding intermediate which was detected as a result of the protection to hydrogen exchange it afforded a limited set of amides; (4) complete the assignments of the sidechain resonances with the goal of determining the solution structure of the protein. %%% The linear amino acid sequence of a protein determines the complex three-dimensional shape that protein acquires in solution. This proposal investigates how that linear information is used to fold the protein into its three-dimensional structure. We will investigate: (1) the role of charge-charge interactions in stabilizing the folded state, (2) the rigidity of the folded state, (3) the time course of the conversion of a random, unfolded polymer chain into the final folded state, and (4) the structure, in solution, of the final folded state.

本建议的目的是为了更好地了解 蛋白质结构、稳定性和动力学的相互关系 以T4溶菌酶为模型。 这种中等大小的蛋白质164 残基具有已知的三维晶体结构。 的 蛋白质具有两个结构域并且没有天然二硫键。 许多突变体存在，调节其热力学 稳定性和超过100种突变晶体结构， 由布莱恩马修斯的实验室在大学的 俄勒冈州。 我们已经指定了野生型的酰胺共振， 利用现代核磁共振技术 共振技术（NMR）。 我们建议使用这些方法， 研究（1）19个羧基的电离行为 目的是为了理解静电， 蛋白质中的极性效应;（2）折叠的动力学 （3）确定在折叠途径中的作用， 折叠中间体，其被检测为 保护氢交换，它提供了一组有限的 酰胺;（4）完成侧链共振的归属 目标是确定 蛋白 %%% 蛋白质的线性氨基酸序列决定了蛋白质的结构。 蛋白质获得的复杂三维形状， 溶液 这个提议调查了线性信息 用来将蛋白质折叠成三维结构。 我们将研究：（1）电荷-电荷相互作用在 稳定折叠状态，（2）折叠状态的刚性， (3)一个随机的， 将未折叠的聚合物链转化为最终折叠状态，和（4）将未折叠的聚合物链转化为最终折叠状态， 在溶液中，最终折叠状态的结构。