摘要
目的寻找与Bat3结合的蛋白质。方法采用串联亲和纯化技术,在Bat3编码序列羧基端融合Strep2和FLAG 2个分子标签,以此为诱饵筛选与其特异结合的蛋白质,最后进行质谱鉴定。结果筛选到1种蛋白质分子,命名为Ubl4A,免疫共沉淀结果显示Ubl4A可与Bat3相互结合。结论成功建立了串联亲和纯化技术平台,分离出可与Bat3相互作用的蛋白质Ubl4A。
Objective To identify the specific protein interactions involved in Bat3-mediated apopto- sis. Methods Tandem affinity purification （TAP） was utilized to investigate Bat3-protein interactions, during which full-length human Bat3 fused with Strep2 and FLAG tag as a bait was used to screen the specific protein- protein interactions. The isolated proteins were identified with mass spectrometry. Results TAP studies showed that Ubl4A was identified as a Bat3-binding partner. Further investigation using co-immunoprecipitation confirmed that Bat3 was associated with Ubl4A. Conclusion TAP was successfully established and is suitable for isolating the binding partners of Bat3.
出处
《中国医学科学院学报》
CAS
CSCD
北大核心
2014年第1期1-4,共4页
Acta Academiae Medicinae Sinicae
基金
国家重大研究计划项目(2011CB944302)
国家重点实验室专项基金(2060204)~~
关键词
Bat3
凋亡
串联亲和纯化
Bat3
apoptosis
tandem affinity purification
作者简介
通信作者：王琳芳电话：010-69156418，电子邮件：wang．1infang@imicams．ac．cn
