2014年第41卷第8期目录
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封面故事:脑疾病造成的社会危害日益严重,且多数无药可医.微量元素硒在预防和延缓此类疾病的过程中具有明显的效果.硒在体内主要是通过硒蛋白发挥生物功能,SelW是人脑中一种重要的硒蛋白,缺硒的条件下,SelW在脑中优先储备,但具体机制和SelW脑中的功能尚不清楚.刘琼研究组以 SelW 突变体 SelW’为“诱饵”,采用酵母双杂交系统筛选人胎脑文库,获得一种与SelW 相互作用的蛋白鞘脂激活蛋白原(PSAP).采用荧光共振能量转移、Pull-down、免疫共沉淀三种方法在体内外分别验证了两者之间的相互作用.结合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,这也为硒在相关领域的研究提供了新的思路和方向.
(陈 平,刘 晴,马孝杰,王诗捷,刘 琼,倪嘉缵. 人脑中硒蛋白W与鞘脂激活蛋白原相互作用的筛选与验证,本期第755~764页)
Cover Story: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. coli. 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.
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综述与专论
研究报告
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