Characterization of the excited state of prion protein using high-pressure NMR technique

本研究的目的是阐明感染机制的构象基础，并表征导致一系列朊病毒疾病的朊病毒蛋白的激发态，这些疾病由朊病毒假说定义。建立了15N标记仓鼠全长朊蛋白的高效表达和纯化方法。在期望能够定量分析在压力下稳定的激发态的NMR横向弛豫色散实验中，尚未获得典型的CPMG分布，并且未能优化这些实验条件。这些结果表明，基本折叠状态和激发态之间的构象波动haPrPC可能会发生超出检测范围（毫秒时间尺度）的NMR R2色散测量。

The purpose of this study is to clarify the conformational basis of infection mechanism and to characterize the excited state of prion protein which causes a series of prion diseases defined by prion hypothesis. We established the protocols for the high-level expression and the purification of 15N-labeled full length hamster prion. In the experiment of NMR transverse relaxation dispersion that expected to enable the quantitative analysis of the excited state which is stabilized under pressure, the typical CPMG profiles have not been obtained and those experimental conditions have failed to be optimized. These results suggest that conformational fluctuations between the basic folded state and the excited state for haPrPC may take place beyond detection range (milli-second time scale) of NMR R2 dispersion measurement.