中文摘要：

硒蛋白W（SelW）是人脑中一种重要的硒蛋白．在缺硒的条件下，SelW在脑中具有优先储备的特性，但其具体机制和在脑中的功能至今尚不清楚．本文以SelW突变体SelW’为“诱饵”，采用酵母双杂交系统对人胎脑文库进行筛选，获得与SelW相互作用的蛋白，其中一种为鞘脂激活蛋白原（PSAP）．采用荧光共振能量转移技术中的受体漂白和敏化发射两种方法，验证了SelW与PSAP的相互作用．构建表达载体在大肠杆菌中成功表达出SelW’，利用Pull．down技术验证了SelW’与PSAP在细胞外的直接相互作用．采用免疫共沉淀的方法验证了上述两种蛋白在昆明小鼠脑组织中的内源性相互作用．基于SelW和PSAP的已知生物功能，推测SelW可能在脑部发育和神经退行性疾病形成过程中发挥着重要作用．

英文摘要：

Selenoprotien W （SelW） is an important selenoprotein that has the priority to be stored in brain when selenium is deficiency. However, the biological function and mechanisms of SelW in brain remain unclear. In this study, human SelW gene was cloned, site-directedly mutated and inserted into the ＂bait＂ plasmid. An interactive protein of SelW, prosaposin （PSAP）, was discovered by screening the human fetal brain cDNA library using the yeast two-hybrid system. To verify the protein-protein interaction, two FRET methods, sensitized emission and acceptor bleaching, were performed respectively in HEK293T cells. Both assays confirmed the interaction between SelW＇ and PSAP. Then the expression vector of SelW＇ was constructed and SelW was overexpressed in E. coll. Pull-down assay was carried out by using the purified SelW＇, and exogenously direct interaction between SelW＇ and PSAP was also verified. Finally, co-IP method was applied to successfully verify the endogenous interaction between the two proteins in the brain tissues of Kunming mice. All these results show that SelW interacts with PSAP directly both in vivo and in vitro. SelW may play a key role in the brain development and neurodegenerative disease formation by interacting with PASP.