猪δ冠状病毒核衣壳蛋白互作的蛋白鉴定及分析

Identification and Analyses of the Proteins Interacting with the Nucleocapsid Protein of the Porcine Deltacoronavirus

初步筛选与鉴定PDCoV N蛋白的宿主互作蛋白。利用免疫共沉淀(Co-immunoprecipitation,CO-IP)和LCMS/MS质谱技术筛选出PDCoV N蛋白的宿主互作蛋白,并进行生物信息学分析,再利用免疫共沉淀进行鉴定。通过免疫共沉淀产物的SDS-PAGE分析发现在40 kD和100 kD左右有明显的差异蛋白条带,通过质谱筛选到了68个与PDCoV N蛋白互作的宿主蛋白,并做了GO、COG和KEGG注释,挑选出2个候选互作蛋白,经过免疫共沉淀验证,PDCoV N蛋白与TUBB2B存在互作关系。该研究为进一步探索PDCoV N蛋白在病毒感染细胞时的作用提供了新方向。

Preliminary screening and identification of the host proteins interacting with the nucleocapsid(N)protein of the porcine deltacoronavirus(DCoV).Co⁃immunoprecipitation and liquid chromatography-tandem mass spectrometry were used to screen out the host proteins interacting with the N protein of the porcine DCoV.Bioinformatics analysis was carried out,and then co⁃immunoprecipitation was used for identification.Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the immunoprecipitation products revealed different protein bands around 40 kDa and 100 kDa.Sixty⁃eight host proteins interacting with the N protein of the porcine DCoV were screened by mass spectrometry.Two candidate interacting proteins(ANXA2 and TUBB2B)were selected by analyses using the Gene Ontology,Clusters of Orthologous Groups,and Kyoto Encyclopedia of Genes and Genomes databases.After co⁃immunoprecipitation verification,the N protein of the porcine DCoV was found to interact with TUBB2B.Our study provides a new direction for further exploration of the role of the N protein of the porcine DCoV in infection.