Study of the molecular mechanisms of abnormal prion protein (PrP) production by exploration of PrP-interacting molecules.

朊病毒疾病是传染性的、致命的神经退行性疾病。朊病毒疾病的标志之一是蛋白酶敏感朊病毒蛋白（PrP^C）的错误折叠和蛋白酶抗性异构体（PrP^<Sc>）的逐渐积累，PrP^C是附着在膜上脂筏上的宿主编码糖蛋白。PrPC向PrPSc的转化似乎发生在细胞表面或沿内吞途径，然而，这种转化的分子基础尚不清楚，特别是关于其他细胞因子在朊病毒复制中的作用。在这项研究中，为了了解转化过程中的分子事件，我们利用一种新建立的细胞表面分子生化标记方法——酶介导的自由基源激活（EMARS），分析了在脂筏富集的抗洗涤剂微域（DRM）中，哪些分子与PrP^C相互作用。在N2a细胞中，我们发现神经元微管调节磷酸化蛋白stthatmin2与PrP共定位。

Prion diseases are transmissible, fatal, neurodegenerative disorders. One of the hallmarks of prion diseases is progressive accumulation of a misfolded and protease-resistant isoform (PrP^<Sc>) of protease-sensitive prion protein (PrP^C) which is a host-encoded glycoprotein attached to the lipid-raft on the membrane. The conversion of PrPC into PrPSc seems to take place at the cell surface or along the endocytic pathway, however, the molecular basis of the conversion remains unclear, particularly regarding role of other cellular factors in prion replication.In this study, to understand the molecular events in the conversion, we analyzed what molecules interact with PrP^C in lipid-raft enriched detergent resistant microdomains (DRM) using the enzyme-mediated activation of radical sources (EMARS), a newly established biochemical labeling method for cell surface molecules. We demonstrated that neuronal microtubule regulatoryphosphoprotein, Sthatmin2 was colocalized with PrP in N2a cells.