STUDY OF PROTEIN DYNAMICS UNDER HIGH HYDROSTATIC PRESSURE

这个子项目是利用资源的许多研究子项目之一。 由NIH/NCRR资助的中心拨款提供。对子项目的主要支持 子项目的首席调查员可能是由其他来源提供的， 包括美国国立卫生研究院的其他来源。为子项目列出的总成本可能 表示该子项目使用的中心基础设施的估计数量， 不是由NCRR赠款提供给次级项目或次级项目工作人员的直接资金。 Acert与韦恩·哈贝尔有着长期的合作，在包括蛋白质动力学在内的分子动力学研究方面的共同兴趣推动了合作。Wayne Hubbell最近已经证明，施加高达2kbar的高静水压力可以填充更高的能量构象，从而使人们能够检测到基态和激发态之间的动态交换，这被强烈地认为是起作用的。Hubbell已经证明，这些状态可以用连续波ESR和脉冲饱和ESR来探测。由于这些状态可以在微秒的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。这为二维Eldor实现其在分子动力学研究中的潜力提供了一个很好的机会。我们将首先对两种自旋标记的蛋白质进行研究：肌红蛋白和IFABP，这是哈贝尔的实验室已经生产出来的。应该指出的是，2D Eldor高压方法的发展具有更广泛的意义，例如，通过提供更多的工具来研究2D-Eldor蛋白质折叠，无论是在有或没有变性剂的稳定状态下，还是在压力跳跃后的动力学研究中。此外，高压下的2D-Eldor可以在Acert的多频率环境下进行，这将有助于更有信心地确定动态变量

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