基于Label-free技术分析牛卵泡蛋白质组分及关键调控蛋白

doi:10.11843/j.issn.0366-6964.2019.05.009

畜牧兽医学报 ›› 2019, Vol. 50 ›› Issue (5): 983-992.doi: 10.11843/j.issn.0366-6964.2019.05.009

基于Label-free技术分析牛卵泡蛋白质组分及关键调控蛋白

郝庆玲¹, 景炅婕², 侯淑宁¹, 许冬梅¹, 赵成萍¹, 朱芷葳¹, 吕丽华^2*, 李鹏飞^1*

1. 山西农业大学生命科学学院, 太谷 030801;
2. 山西农业大学动物科技学院, 太谷 030801

收稿日期:2018-09-27 出版日期:2019-05-23 发布日期:2019-05-23
通讯作者: 李鹏飞,主要从事动物生殖生理方面的研究,E-mail:adamlpf@126.com;吕丽华,主要从事动物生殖生理及繁殖调控生物技术方面的研究,E-mail:lihualvsxau@126.com
作者简介:郝庆玲(1994-),女,山西原平人,硕士生,主要从事动物生殖生理研究,E-mail:1397542322@qq.com;景炅婕(1991-),女,山西沁水人,博士生,主要从事动物生殖生理研究,E-mail:ycjingjiongjie@163.com
基金资助:
国家自然科学基金（31873002）；山西省国际科技合作项目（201603D421006）；山西省三晋学者和人才引进项目；山西省重点研发计划项目（201703D221020-1；201803D31062）；山西省研究生优秀创新项目（2017BY068）；山西农业大学创新基金项目（zdpy201403/201503）；山西农业大学青年拔尖创新人才支持计划（TYIT201403）

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

HAO Qingling¹, JING Jiongjie², HOU Shuning¹, XU Dongmei¹, ZHAO Chengping¹, ZHU Zhiwei¹, LÜ Lihua^2*, LI Pengfei^1*

1. College of Life Science, Shanxi Agricultural University, Taigu 030801, China;
2. College of Animal Science and Veterinary Medicine, Shanxi Agricultural University, Taigu 030801, China

Received:2018-09-27 Online:2019-05-23 Published:2019-05-23
Supported by:

摘要/Abstract

摘要：

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。

Abstract:

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.

中图分类号:

S823.2

郝庆玲, 景炅婕, 侯淑宁, 许冬梅, 赵成萍, 朱芷葳, 吕丽华, 李鹏飞. 基于Label-free技术分析牛卵泡蛋白质组分及关键调控蛋白[J]. 畜牧兽医学报, 2019, 50(5): 983-992.

HAO Qingling, JING Jiongjie, HOU Shuning, XU Dongmei, ZHAO Chengping, ZHU Zhiwei, Lü Lihua, LI Pengfei. Proteomic Analysis of Follicular Components and Key Regulatory Proteins in Bovine Based on Label-free Technology[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(5): 983-992.

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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

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