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

Pron病是一种传染性、致命性、神经退行性疾病。蛋白水解酶敏感的蛋白(PrP^C)是一种宿主编码的糖蛋白，附着在膜上的脂筏上。PrPC向PrPSc的转化似乎发生在细胞表面或沿胞内途径，但其分子基础尚不清楚，尤其是其他细胞因子在PrP复制中的作用。在本研究中，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的生化标记方法--酶介导的自由基源活化(EMARS)，分析了在脂筏富含洗涤剂的微区(DRM)中PrP^C与PrP^C相互作用的分子。我们在N2a细胞中证实了神经元微管调节蛋白Sthen2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.