神经病靶标酯酶调控结构域结合蛋白筛选及其在COS-7细胞中的表达

Hunting for novel proteins in neuropathy target esterase regulatory domain with yeast two-hybrid system

本研究采用酵母双杂交系统探寻与神经病靶标酯酶(NTE)调控结构域相互作用的蛋白因子,揭示与NTE信号转导相关的可能机制。通过构建含有NTE调控结构域的诱饵蛋白载体筛选胎脑文库,并将筛选得到的阳性克隆在酵母中进行了验证,随后在哺乳动物细胞中表达了该蛋白。生物信息学分析显示:该阳性克隆为前列腺素受体结合蛋白54(ARA54),具有泛素连接酶活性,提示细胞可能存在依赖于细胞周期的NTE活性调节机制,为阐明NTE生理功能创造了条件[动物学报51(5):840-844,2005]。

To explore protein factors that may interact with regulatory domain of neuropathy target esterase （NTE） in signal transduction pathways, the bait plasmid of NTE-regulatory domain eDNA was constructed and screened in human fetal brain eDNA library by the method of yeast two-hybrid. The candidate positive clones were retested in yeast, and its expression was verified in mammalian cells. The results of bioinformatics showed that androgen receptor-associated protein 54 （ARA54）, a ubiquitin protein ligase, may be an essential controller of the NTE in the process of cell cycle. This result will put great opportunity for understanding the physiological mechanism of the NTE [Acta Zoologica Sinica 51 （5）： 840-844, 2005].