International Research Fellowship Program: Invisible, Excited State Structures from Relaxation Dispersion NMR Spectroscopy

国际研究奖学金计划：弛豫色散核磁共振波谱中的不可见激发态结构

• 批准号: 0853108
• 负责人: Michael Latham
• 金额: $ 12.6万
• 依托单位: Latham Michael P
• 依托单位国家: 美国
• 项目类别: Fellowship Award
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-10-01 至 2011-09-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0853108&HistoricalAwards=false
• 关键词: International; Research; Fellowship; Program; Invisible

0853108LathamThis award is funded under the American Recovery and Reinvestment Act of 2009 (Public Law 111-5).The International Research Fellowship Program enables U.S. scientists and engineers to conduct nine to twenty-four months of research abroad. The program's awards provide opportunities for joint research, and the use of unique or complementary facilities, expertise and experimental conditions abroad.This award will support a twenty-four-month research fellowship by Dr. Michael P. Latham to work with Dr. Lewis E. Kay at the University of Toronto in Canada.Determining models for biomolecules is a primary goal in structural biology, and these models can be used to gain insight into biomolecular function. Generally, high-resolution models of biomolecules are only obtainable for low energy, ground state conformations. While these static ground state structures offer a wealth of information, the dynamic nature of biomolecules must also be investigated to more fully understand the relationship between structure, function and mechanism. Thus, protein function may not be fully appreciated without an understanding for the excursion of ground state structures to higher energy excited states. However, structural biology studies of excited states are complicated by the low populations and transient nature of these often ?invisible? excited states. NMR spectroscopy is uniquely capable of site-specifically probing protein dynamics over a wide range of timescales. The powerful NMR relaxation dispersion technique is particularly sensitive to the presence of lowly populated excited states. The objective of this proposal is to develop and apply these techniques to structural studies of 'invisible' excited states in the folding pathway of two modular domains. Specifically, new relaxation dispersion methods are being developed, which complement other recently described techniques, to probe the structures of excited states using NMR chemical shift values and residual dipolar couplings. These methods are then extended to generate models for low energy, on-pathway folding intermediates of the Fyn SH3 and human HYPA/FBP11 FF domains, which will represent some of the first pictures of folding intermediates and ?invisible? excited states. While the techniques utilized here are initially applied to protein folding, they should be generally applicable in the study of enzyme catalysis and protein ligand binding. Moreover, there is an increased appreciation for the role of protein misfolding in disease states, such as cystic fibrosis, Alzheimer?s disease and diseases resulting from prions. As the case with cystic fibrosis, some genetic diseases are intimately coupled with protein misfolding, resulting in the formation of stable, inactive intermediate states. Thus, this study could open the door to other structural studies of 'invisible' intermediate states involved in protein folding/misfolding associated with disease states.

[0853108] latham该奖项是根据2009年美国复苏和再投资法案（公法111-5）资助的。国际研究奖学金项目使美国科学家和工程师能够在国外进行9至24个月的研究。该计划的奖励为联合研究提供了机会，并利用独特或互补的设施、专业知识和国外的实验条件。该奖项将支持Michael P. Latham博士与加拿大多伦多大学Lewis E. Kay博士一起进行为期24个月的研究。确定生物分子的模型是结构生物学的主要目标，这些模型可以用来深入了解生物分子的功能。一般来说，生物分子的高分辨率模型只能用于低能基态构象。虽然这些静态基态结构提供了丰富的信息，但生物分子的动态性质也必须进行研究，以更充分地了解结构、功能和机制之间的关系。因此，如果不了解基态结构向高能激发态的偏移，蛋白质的功能可能无法得到充分的认识。然而，激发态的结构生物学研究由于这些通常是“看不见的”的低种群和短暂性而变得复杂。激发态。核磁共振波谱是唯一能够在广泛的时间尺度上特异性探测蛋白质动力学的位点。强大的核磁共振弛豫色散技术对低密度激发态的存在特别敏感。本提案的目的是发展和应用这些技术在两个模域折叠路径中“不可见”激发态的结构研究。具体来说，新的弛豫色散方法正在开发中，它补充了其他最近描述的技术，利用核磁共振化学位移值和剩余偶极耦合来探测激发态的结构。然后将这些方法扩展到生成Fyn SH3和人类HYPA/FBP11 FF结构域的低能量，通路上折叠中间体的模型，这将代表折叠中间体和“不可见”的一些第一张图片。激发态。虽然这里使用的技术最初应用于蛋白质折叠，但它们应该普遍适用于酶催化和蛋白质配体结合的研究。此外，人们越来越认识到蛋白质错误折叠在疾病状态中的作用，如囊性纤维化、阿尔茨海默病(Alzheimer？S疾病和由朊病毒引起的疾病。如囊性纤维化，一些遗传疾病与蛋白质错误折叠密切相关，导致形成稳定的、无活性的中间状态。因此，这项研究可以为与疾病状态相关的蛋白质折叠/错误折叠相关的“不可见”中间状态的其他结构研究打开大门。