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

使用高压核磁共振技术表征朊病毒蛋白的激发态

• 批准号: 22770158
• 负责人: MAENO Akihiro
• 金额: $ 2.58万
• 依托单位: Kinki University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22770158/
• 关键词: プリオン蛋白質; 核磁気共鳴; 圧力; 励起構造

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.

本研究的目的是阐明朊病毒感染机制的构象基础，并表征由朊病毒假说定义的一系列朊病毒疾病的朊病毒蛋白的激发态。建立了15n标记全长仓鼠朊病毒的高水平表达和纯化方案。在期望定量分析压力稳定激发态的核磁共振横向弛豫色散实验中，没有得到典型的CPMG谱线，实验条件也没有优化。这些结果表明，在核磁共振R2色散测量的探测范围（毫秒时间尺度）之外，haPrPC的基本折叠态和激发态之间可能发生构象波动。

项目成果

Cavity Hydration : A Gate Way to the High-Energy Paradigm?

腔内水合：通向高能量范式的大门？

• 发表时间: 2010
• 作者: A.Nakatsuma;T.Yamashita;K.Sasaki;A.Kawanabe;K.Inoue;Y.Furutani;Y.Shichida;H.Kandori;牧野正知;前野覚大
• 通讯作者: 前野覚大

Cavity hydration and the dynamics of a protein

腔内水合和蛋白质的动力学

• 发表时间: 2011
• 作者: 前野覚大;北原亮;鎌足雄司;Sunilkumar PN;赤坂一之
• 通讯作者: 赤坂一之