基于Label-free技术分析牛卵泡蛋白质组分及关键调控蛋白

Proteomic Analysis of Follicular Components and Key Regulatory Proteins in Bovine Based on Label-free Technology

旨在研究牛卵泡发育过程中蛋白质组表达变化并筛选卵泡发育关键调控蛋白,利用非标记(label-free)定量蛋白质组学技术对牛卵泡颗粒细胞(granulesa cells, GCs)蛋白质组分进行比较分析。采集牛发情周期优势卵泡(dominant follicles, DF)和从属卵泡(subordinate follicles, SF),分别分离GCs,并提取总蛋白,胰蛋白酶酶解,液相色谱-串联质谱(LC-MS/MS)进行蛋白质组分分析,数据库检索分析DF和SF中蛋白质表达情况,并应用生物信息学方法筛选牛卵泡发育关键调控蛋白。结果表明:本试验从30 321个肽段中共成功鉴定出3 409种蛋白质(FDR≤0.01),其中,DF中表达2 895种蛋白质,SF中表达3 102种蛋白质,获得差异表达蛋白质(差异倍数>2,P<0.05) 259种,17种差异表达蛋白质可能与牛卵泡优势化过程相关,SERPINB2可能调控牛卵泡闭锁。该研究筛选获得的牛卵泡差异表达蛋白质和特异表达蛋白质,丰富了牛卵泡发育调控理论,为进一步研究卵泡闭锁及优势化奠定基础。

The purpose of this study was to investigate changes in proteome expression and screen key regulatory proteins during bovine follicular development,and protein components of GCs(granulesa cells)were analyzed by label-free quantitative proteomics technology.DF(dominant follicles)and SF(subordinate follicles)during the bovine estrous cycle were collected,GCs were separated,total proteins were extracted,proteins were digested with trypsin and analyzed by LC-MS/MS,the expression of proteins in DF and SF were analyzed after database retrieval,and key regulatory proteins were screened by bioinformatics method.In this study,3 409 proteins were successfully identified from 30 321 peptide fragments (FDR≤0.01),of which 2 895 were DF proteins and 3 102 were SF proteins,259 differentially expressed proteins were identified (Fold change>2,P <0.05),17 differential expressed proteins might be related to the process of bovine follicular dominance,and SERPINB2 might regulate bovine follicular atresia.The results enriched the studies on DF and SF proteome and laid a foundation for further studies on follicular atresia and dominance theory by identify differential expressed proteins and specific expression proteins.