STUDY OF PROTEIN DYNAMICS UNDER HIGH HYDROSTATIC PRESSURE

高静水压下蛋白质动力学的研究

• 批准号: 8364112
• 负责人: PETER P BORBAT
• 金额: $ 0.37万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-01 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8364112
• 关键词: Collaborations; Development; Frequencies; Funding; Grant; Hydrostatic Pressure; Laboratories; Methods; Myoglobin; National Center for Research Resources; Physiologic pulse; Principal Investigator; Protein Dynamics; Proteins; Research; Research Infrastructure; Resources; Source; Spin Labels; Technology; Time; United States National Institutes of Health; conformer; cost; interest; molecular dynamics; pressure; protein folding; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. ACERT has a long-standing collaboration with Wayne Hubbell, fueled by common interests in the study of molecular dynamics including protein dynamics. It has been shown recently by Wayne Hubbell that application of high hydrostatic pressure of up to 2 kbar populates higher energy conformers, and thereby enables one to detect dynamic exchange between the ground state and excited state, which is strongly believed to be functional. Hubbell has shown that these states can be probed by CW ESR and pulse saturation ESR. Since these states can undergo exchange on a microsecond time-scale, it would be logical to apply 2D ELDOR to study protein dynamics and conformational exchange in proteins under high pressure. This presents a great opportunity for 2D ELDOR to realize its potential in studying molecular dynamics. We will conduct the study initially on two spin-labeled proteins: Myoglobin and IFABP, which Hubbell's laboratory has already produced. It should be noted that the development of a 2D ELDOR high pressure method has broader implications, for example, by providing more tools to study by 2D-ELDOR protein folding, both in the steady state with or without denaturant and kinetically after a pressure jump. Furthermore, 2D-ELDOR at high pressure can be conducted at ACERT in a multi-frequency context, which would help to determine dynamic variables with greater confidence

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的首席研究员可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 ACERT 与 Wayne Hubbell 有着长期的合作关系，这得益于对分子动力学（包括蛋白质动力学）研究的共同兴趣。 Wayne Hubbell 最近表明，应用高达 2 kbar 的高静水压力会填充更高能量的构象异构体，从而使人们能够检测基态和激发态之间的动态交换，这被强烈认为是功能性的。 Hubbell 表明，这些状态可以通过 CW ESR 和脉冲饱和 ESR 来探测。 由于这些状态可以在微秒时间尺度上进行交换，因此应用 2D ELDOR 来研究高压下蛋白质的动力学和构象交换是合乎逻辑的。 这为 2D ELDOR 提供了一个发挥其在分子动力学研究潜力的绝佳机会。我们将首先对两种自旋标记蛋白进行研究：肌红蛋白和 IFABP，Hubbell 实验室已经生产了这两种蛋白。值得注意的是，2D ELDOR 高压方法的发展具有更广泛的意义，例如，通过提供更多工具来研究 2D-ELDOR 蛋白质折叠，无论是在有或没有变性剂的稳态下，还是在压力跳跃后的动力学上。此外，高压下的 2D-ELDOR 可以在多频率背景下在 ACERT 上进行，这将有助于更有信心地确定动态变量