HIGH PRESSURE NMR STUDIES OF PROTEINS

蛋白质的高压核磁共振研究

• 批准号: 2181382
• 负责人: JIRI JONAS
• 金额: $ 20.86万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-07-01 至 1998-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2181382
• 关键词: biomedical equipment development; biophysics; chemical kinetics; chemical stability; circular dichroism; conformation; fatty acid binding protein; hydropathy; lasers; lysozyme; mechanical pressure; molecular dynamics; mutant; myoglobin; nuclear magnetic resonance spectroscopy; nuclease; pancreatic ribonuclease; photochemistry; protein denaturation; protein folding; protein structure; temperature; thermodynamics

The long-term objective of the proposed research is to contribute to the understanding of the stability, folding intermediates, and dynamics of proteins by using pressure as the fundamental thermodynamic variable. Unique high pressure equipment, suitable for measurements from 0.1 MPa (l bar) to l GPa (10 kbar) and from -25 degrees C to lOO degrees C, will be used in conjunction with a variety of l-D and 2-D high resolution NMR techniques to study the effects of pressure on the unfolding/folding equilibria, conformational states of native and compact denatured states, and the dynamic properties of several model proteins. In addition, the cold-denaturation behavior of these proteins will be studied by taking advantage of the depression of the freezing point of water by pressure. The pressure, cold, heat, and chemically denatured states of the model proteins will be examined and compared, with special emphasis on the compact denatured states observed by cold and pressure denaturation. The model proteins selected for these studies include hen egg-white lysozyme, bacteriophage T4 lysozyme, horse myoglobin, sperm-whale myoglobin, staphylococcal nuclease, and bovine pancreatic ribonuclease A, as well as selected mutants of T4 lysozyme, sperm-whale myoglobin, staphylococcal nuclease, and bovine pancreatic ribonuclease A. To complement the NMR measurements at high pressure, especially on the compact denatured states of the model proteins and their mutants, circular dichroism (CD) and laser-photo chemically induced dynamic nuclear polarization (CIDNP) experiments will be performed. High-pressure CD cells and CIDNP probes will be designed and constructed.

拟议研究的长期目标是为 了解稳定性，折叠中间体和动态 通过使用压力作为基本热力学变量，蛋白质。 独特的高压设备，适用于0.1 MPa的测量 bar）至L GPA（10 kbar）和-25度至较高的c 与各种L-D和2-D高分辨率NMR结合使用 研究压力对展开/折叠的影响的技术 平衡，天然和紧凑型状态的构象状态， 以及几种模型蛋白的动态特性。 另外， 将研究这些蛋白质的冷饱和行为 压力降低冰点的优势。 模型的压力，冷，热和化学变性状态 将检查和比较蛋白质，并特别强调 紧凑的变性状态通过冷和压变性观察到。这 选择这些研究的模型蛋白质包括鸡蛋白溶菌酶， 噬菌体T4溶菌酶，马肌红蛋白，精子 - 鞭毛肌红蛋白， 葡萄球菌核酸酶和牛胰腺核糖核酸酶A，以及 选定的T4溶菌酶，精子旋转肌红蛋白，葡萄球菌的突变体 核酸酶和牛胰腺核糖核酸酶A. 补充高压下的NMR测量值，尤其是在 紧凑的模型蛋白质及其突变体的变性状态，圆形 二分（CD）和激光化学诱导的动态核 将进行极化（CIDNP）实验。 高压CD 细胞和CIDNP探针将设计和构造。