酵母双杂交方法筛选与NGF受体TrkA相互作用的新蛋白质  

Screening for new protein binding with nerve growth factor receptor TrkA using yeast two-hybrid method

作　　者：张勇[1] 黄爱军[1] 曹莉[1] 阎志勇[1] 刘秀杰[1] 张骐[1] 路长林[1] 何成[1] 

机构地区：[1]第二军医大学基础医学部神经生物学教研室,上海200433

出　　处：《第二军医大学学报》2002年第6期586-589,共4页Academic Journal of Second Military Medical University

基　　金：上海市曙光计划;上海市科学技术发展基金资助项目(0 1JC14 0 0 4) ;军队医学杰出人才基金资助项目 (0 1J0 11) ;校科研联合攻关资助项目 (2 0 0 1) .

摘　　要：目的 :寻找新的 Trk A可能底物或调控蛋白 ,以深入认识 Trk A下游信号转导机制。方法 :将 Trk A胞内域与 L ex A蛋白融合作为诱饵蛋白 ,应用酵母双杂交方法筛选人脑 L ex A c DNA文库 ,经β-半乳糖苷酶活性鉴定和测序分析进一步鉴别 ,获得阳性克隆。 结果 :明确阳性克隆中的 Trk AIC- BP1为一种新的 Trk A结合蛋白。免疫共沉淀实验证实了 Trk AIC- BP1和Trk A在酵母中的相互作用。结论 :首次筛选到 Trk A结合蛋白—— Trk AIC- BP1,它可能在 TrkObjective:To search for the possible intracellular substrates or regulatory proteins of the nerve growth factor(NGF) receptor TrkA, as to understand the mechanisms of receptor downstream signal transduction.Methods: The TrkA IC (intracellular part of TrkA) was fused to LexA, the product was used as a bait to screen a human brain LexA two hybrid cDNA library and 3 positive clones were obtained. Results: Among them TrkA IC BP1 was identified as a new TrkA binding protein by β galactosidase activity and sequence analysis. The interaction between the TrkA IC BP1 and the TrkA IC was confirmed by immunoprecipitation in yeast. Conclusion: TrkA IC BP1 is firstly found as TrkA binding protein by yeast two hybrid screening, and it may be important to the regulation of TrkA mediated neuronal differentiation.

关 键 词：酵母杂交方法 NGF受体 TRKA 相互作用 蛋白质 TrkA^IC-BP1 

分 类 号：Q513[生物学—生物化学] Q257