SAXS-DETECTED DYNAMICS OF FAST-FOLDING PROTEINS

SAXS 检测快速折叠蛋白质的动力学

• 批准号: 7601757
• 负责人: MARTIN GRUEBELE
• 金额: $ 3.53万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-01 至 2008-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7601757
• 关键词: Cell Nucleus; Circular Dichroism Spectroscopy; Computer Retrieval of Information on Scientific Projects Database; Dependence; Funding; Goals; Grant; Hydrophobic Interactions; Institution; Kinetics; Maps; Measurement; Measures; Phase; Proteins; Publications; Radiation; Rate; Research; Research Personnel; Resources; Roentgen Rays; Science; Source; Temperature; Testing; Time; United States National Institutes of Health; data modeling; follow-up; lambda repressor; mutant; protein aggregation; protein folding; radius bone structure; size

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This ongoing project follows up on a previous BioCAT project to study fast folding dynamics of lambda repressor by a combination of small angle X-ray scattering and circular dichroism spectroscopy (Dumont et al., Protein Science 15, 2596-2604 (2006)). We are examining the aggregation of lambda repressor and of fyn SH3 to obtain the potential of mean force between molecules and hence the size of the critical aggregation nucleus. The critical nucleus is an important measure of protein aggregation, but difficult to determine. We are also studying the folding dynamics of a downhill-folding mutant of lambda repressor by stopped-flow small angle X-ray scattering. The goal is to test the hypothesis that loss of the hydrophobic interaction eventually leads to a "turn-around" in the folding rate. We recently had our first beam time for this project, with the next planned time coming up in February 2007. We have completed radiation damage test on the proteins, all the concentration studies needed for the aggregation study (which is now in a phase of modeling the data to prepare for publication), and some of the kinetics measurements for the downhill lambda repressor mutants. More kinetics, including measurements at different final denaturant concentrations will be taken for both proteins next, to provide a complete map of the temperature dependence and time dependence of the radius of gyration.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 这个正在进行的项目跟进了一个先前的BioCat项目，该项目通过小角度X射线散射和圆形二分法光谱的结合来研究Lambda阻遏物的快速折叠动力学（Dumont等人，蛋白质科学15，2596-2604（2006））。 我们正在研究Lambda阻遏物和FYN SH3的聚集，以获得分子之间的平均力潜力，并因此获得了临界聚集核的大小。 临界核是蛋白质聚集的重要度量，但难以确定。 我们还通过停止流动的小角度X射线散射来研究Lambda阻遏物下坡折叠突变体的折叠动力学。 目的是检验以下假设：疏水相互作用的丧失最终导致折叠率的“转弯”。 最近，我们在2007年2月出现了下一个计划的时间。我们已经完成了蛋白质的辐射损伤测试，集合研究所需的所有集中研究（现在正处于建模数据的阶段，以准备出版物准备出版物），以及一些动力学测量值，用于下落Lambda Repressor repressor Mutants。 接下来，将对两种蛋白质进行更多的动力学，包括在不同最终变性浓度下的测量，以提供回旋半径半径的温度依赖性和时间依赖性的完整图。