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Pron蛋白的激发状态，它导致了Pron假说所定义的一系列Pron疾病。我们建立了高效表达和纯化15N标记的全长仓鼠蛋白的方法。在核磁共振横向弛豫色散实验中，为了能够对压力下稳定的激发态进行定量分析，没有得到典型的CPMG曲线，这些实验条件也没有得到优化。这些结果表明，haPrPC的基本折叠态和激发态之间的构象波动可能发生在核磁共振R2色散测量的探测范围(毫秒时间尺度)之外。

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.