中国农业科学 ›› 2020, Vol. 53 ›› Issue (14): 2956-5963.doi: 10.3864/j.issn.0578-1752.2020.14.018

• 畜牧·兽医·资源昆虫 • 上一篇    下一篇

基于绵羊胚胎骨骼肌蛋白质组学的PI3K-AKT信号通路分析

王欣悦(),石田培,赵志达,胡文萍,尚明玉,张莉()   

  1. 中国农业科学院北京畜牧兽医研究所,北京 100193
  • 收稿日期:2019-08-29 接受日期:2020-03-30 出版日期:2020-07-16 发布日期:2020-08-10
  • 通讯作者: 张莉
  • 作者简介:王欣悦,E-mail:wxyanimalgenetic@163.com
  • 基金资助:
    国家自然基金联合基金重点支持项目(U1503285);中国农业科学院基本科研业务费重大项目储备计划(Y2017XM02)

The Analysis of PI3K-AKT Signal Pathway Based on the Proteomic Results of Sheep Embryonic Skeletal Muscle

WANG XinYue(),SHI TianPei,ZHAO ZhiDa,HU WenPing,SHANG MingYu,ZHANG Li()   

  1. Institute of Animal Sciences, Chinese Academy of Agriculture Sciences, Beijing 100193
  • Received:2019-08-29 Accepted:2020-03-30 Online:2020-07-16 Published:2020-08-10
  • Contact: Li ZHANG

摘要:

【目的】绵羊是重要的经济动物,其骨骼肌生长发育与产肉性能密切相关。胚胎期是绵羊骨骼肌生长发育的关键阶段,挖掘分析绵羊胚胎骨骼肌蛋白质组数据,为揭示绵羊肌肉发育重要时间节点、筛选绵羊胚胎骨骼肌生长发育调控蛋白质提供依据。【方法】本团队已对妊娠第85天、第105天和第135天的中国美利奴绵羊胚胎背最长肌进行串联质谱(tandem mass tag, TMT)蛋白质定量,鉴定到1316种差异丰度蛋白质。现利用GO、KEGG和R等方法对这些差异丰度蛋白质开展聚类、功能注释和通路分析等生物信息学分析。【结果】基于前期研究结果对差异丰度蛋白质进行R语言聚类,分析结果显示,cluster 5类蛋白在胚胎骨骼肌第105天具有较高丰度。对cluster 5 蛋白进行GO和KEGG富集分析发现,该类蛋白质参与胞内蛋白质代谢过程,显著富集于PI3K-AKT信号通路中,而在该信号通路中RAC-β丝氨酸/苏氨酸蛋白激酶X1(AKT2)具有较高表达丰度。蛋白质生物信息学结果表明,AKT2蛋白由481个氨基酸构成,AKT2蛋白理论分子量为55.58kD,由66个带正电荷的氨基酸残基和72个带负电荷的氨基酸残基组成,理论等电点为6.08,亲水性平均系数-0.454,属于亲水性蛋白。预测AKT2蛋白的481个氨基酸全部位于膜外,属于膜受体蛋白。AKT2蛋白有12个N-端糖基化位点,71个磷酸化位点,与蛋白酶K相似度为99%,属于蛋白酶催化亚基家族。【结论】绵羊胚胎骨骼肌蛋白质组数据发现,第105天是绵羊胚胎骨骼肌纤维由增殖分化到增大增粗的转折点,具有调控绵羊胚胎骨骼肌纤维生长发育作用的PI3K-AKT信号通路在该节点显著富集,AKT2是调控该信号通路的重要候选蛋白。综上,本研究结果对揭示胚胎骨骼肌生长发育及其调控分子机制具有重要理论指导意义。

关键词: 绵羊(Ovis aries), 胚胎背最长肌, 蛋白质组学, 生物信息学分析

Abstract:

【Objective】Sheep is an important economic livestock and its skeletal muscle growth and development have a deep bond with meat production traits. The sheep embryonic period is an essential stage for skeletal muscle growth, analyzing and mining the proteome data of sheep embryonic skeletal muscle in this period has a great significance to reveal the muscle development process and screen their key regulation proteins.【Method】The longissimus dorsi of Chinese merino sheep at embryonic age of 85 days, 105 days and 135 days were selected for protein qualification by using tandem mass tag (TMT) and 1316 differential abundance proteins were obtained finally. GO, KEGG and R bioinformatic methods were used to cluster, annotate and analyze the differential abundance proteins. And the candidate proteins were testified by using bioinformatic methods.【Result】Based on the previous results, the cluster analysis on differential abundance proteins illustrated that the cluster 5 proteins were significantly expressed on embryonic age of 105 days with high abundance. GO and KEGG analysis on cluster 5 proteins showed these proteins were significantly involved in protein metabolism biology process and notably enriched in PI3K-AKT signal pathway in which RAC-beta serine/threonine-protein kinase isoform X1(ATK2) has a high abundance. Meanwhile, the results of bioinformatics showed that the AKT2 was composed of 481 amino acids and the theoretical molecular weight was 55.58kD. It consists of 66 positively charged amino acid residues and 72 negatively charged amino acid residues, the theoretical isoelectric point was 6.08, the hydrophilicaverage coefficient was -0.454, 12 N-terminal glycosylation sites and 71 phosphorylation sites were found in AKT2. The homology of AKT2 and protein kinase-like (PK-like) was 99% and it belongs to the family of protein kinases catalytic subunit.【Conclusion】The proteome data analysis of sheep embryonic skeletal muscle showed that embryonic age of 105 days is a key point of sheep embryonic skeletal fiber cell from proliferation and differentiation to hypertrophy. The PI3K-AKT signaling pathway which has function of regulating growth and development of embryonic skeletal muscle fibers was significantly enriched, and ATK2 is a key candidate regulation protein in this pathway. To summarize, the study has a theoretical guiding significance to reveal the growth and development and its molecular regulation mechanism of embryonic skeletal muscle.

Key words: sheep (Ovis aries), embryonic longissimus dorsi, proteomic, bioinformatics analysis