免疫共沉淀联合质谱技术筛选宿主因子CypB的互作蛋白

Identification of the interacting proteins with CypB by co-immunoprecipitation combining with mass spectrometry

为探讨亲环蛋白B(cyclophilin B,CypB)促进羊传染性脓疱病毒(orf virus,ORFV)复制增殖的机制,以ORFV早期感染MDBK细胞为试验材料,利用免疫共沉淀技术筛选与CypB互作的蛋白。免疫共沉淀产物的Western blot和SDS-PAGE的分析结果显示,在大小约为25 000处可见有明显的差异条带。切下该差异蛋白条带进行质谱鉴定分析,将获得的原始质谱数据使用Mascot软件中的串级质谱数据搜索功能(MS/MS Ions Search)进行搜索鉴定,初步筛选获得6个可能与CypB相互作用的病毒相关蛋白。为进一步深入研究CypB在ORFV早期侵染过程中的作用奠定基础。

In order to reveal the mechanism of cyclophilin B （CypB） contributing to ORFV replica- tion,the interacting proteins with CypB are identified by co-immunoprecipitation assay using the MDBK cells infected with ORFV as experimental materials. An obvious protein band was observed at the size of about 25 000 by Western blot and SDS-PAGE analysis. The protein band was cut from the gel and analyzed by mass spectrometry （MS）. The data obtained by MS results were i- dentified by MS/MS Ions Search function of Mascot software. Six proteins related to ORFV were obtained,which might interact with CypB. The above results will establish foundation for further determining the role of CypB in ORFV early infection.