Structural analysis of zoonosis-related high-molecular protein aggregation by double electron-electron resonance (DEER) technique

在本研究中，为了获得PrPs'病理纤维形态的结构信息，我们采用连续波（CW） ESR （x波段）和脉冲ESR （q波段）的定点自旋标记（SDSL）测量了D178N突变的病理纤维形态的分子间距离。在pH4.0中暴露于1M盐酸胍形成纤维结构后，用CW-ESR法测定其ESR光谱。采用连续波ESR （CW-ESR）和四脉冲双电子-电子共振（DEER）技术测量病理纤维形态的旋间距离。分子间距离信息将有助于估计家族性朊病毒疾病的致病纤维结构，因为通过核磁共振或x射线晶体学难以分析纤维蛋白等大分子。

In this study, to obtain the structural information for this pathological fibrous form of PrPs', we measured the intermolecular distance in the pathological fibrous form with D178N mutation by the site-directed spin labeling (SDSL) with continuous wave (CW) ESR (X-band) and pulsed ESR (Q-band). After the formation of the fibrous structure by exposure to 1M of guanidine hydrochloride in pH4.0, the ESR spectra were measured by CW-ESR. The interspin-distances in pathological fibrous form were measured by continuous wave ESR (CW-ESR) and the four-pulse double electron-electron resonance (DEER) technique. The information of intermolecular distances will be useful to estimate the pathogenic fibrous structure of the familial prion disease, since it is difficult to analyze the macromolecules such as the fibrous protein by NMR or X-ray crystallography.