基于Label-free技术比较马和驴血清蛋白质组分差异

doi:10.11843/j.issn.0366-6964.2021.011.011

摘要/Abstract

摘要： 旨在探究马和驴血清蛋白质生物学特征，为马属动物种间生理特征比较和健康保障提供理论依据。本研究选取辽宁省大连市某集约化养殖场的9匹蒙古马和9头辽西驴，雌性，年龄4~10岁，均处于正常发情周期的间情期，健康无病、精神状态及采食状况良好，分别提供相同的饲养条件。采集马和驴各9份血清样品分别随机分成3组，每组内的3份血清样品均匀混合成1个生物学重复，各获得3个生物学重复的血清样品。利用Label-free蛋白质组学技术及生物信息学方法对马和驴的血清蛋白质组分进行比较研究，提取血清蛋白质，再对蛋白质进行酶解，液相色谱-串联质谱（LC-MS/MS）进行蛋白质组分分析，数据库检索分析马和驴血清样品中蛋白质表达情况，并应用生物信息学方法筛选马和驴血清差异关键调控蛋白质。结果显示，本研究共鉴定出361种蛋白质，其中，马血清中表达288种蛋白质，分子量范围为1.52~511.24 ku；驴血清中表达244种蛋白质，分子量范围为1.53~611.47 ku；共获得231种显著差异表达蛋白质（差异倍数≥1.5，P≤0.05）。差异表达蛋白质主要参与蛋白质激活、补体激活、免疫应答、凝血和脂蛋白氧化调控等生物学过程。显著富集的KEGG通路为补体和凝血级联，吞噬体，内质网蛋白质加工，抗原加工递呈，甘氨酸、丝氨酸和苏氨酸的代谢，癌症蛋白多糖等。通过差异蛋白互作分析显示，差异表达蛋白质紧密相连富集最为显著的功能模块主要为代谢途径、补体和凝血级联、吞噬体，HSP90AA1、HSPA8、APOD、APOM、SERPING1、MASP1、CALR、TUBA1B、TUBB等处在关系互作网的重要节点。马和驴在生理条件下的血清蛋白质组成特征存在一定差异，该研究为进一步揭示马属动物种间生理特征差异和有效保障健康提供数据支撑。

关键词: 马, 驴, 血清, Label-free, 蛋白质组

Abstract: The purpose of this study was to explore the biological characteristics of serum proteins in horses and donkeys, and to provide a theoretical basis for the comparison of the physiological characteristics among equine species and health protection. In this study, 9 Mongolian horses and 9 Liaoxi donkeys, female, aged 4-10 years old, were selected from an intensive farm in Dalian city, Liaoning province. They were all in the interestrus of normal estrous cycle. They were healthy, disease-free, mental state and eating well, and provided the same feeding conditions. Nine serum samples of horses and donkeys were collected and randomly divided into 3 groups, respectively. Three serum samples in each group were evenly mixed into one biological repeat, and 3 serum biological repeats were obtained in each group. In this study, Label-free proteomics and bioinformatics methods were used to compare the serum protein components of horses and donkeys,serum proteins were extracted, and then the proteins were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). The expression of proteins in horse and donkey serum samples was analyzed by database search. The different key regulatory proteins in serum between horses and donkeys were screened by bioinformatics method.The results showed that a total of 361 proteins were identified, of which 288 proteins were identified in horse serum with molecular weight from 1.52 to 511.24 ku, and 244 proteins were identified in donkey serum with molecular weight from 1.53 to 611.47 ku, and 231 differentially expressed proteins were obtained (fold change≥1.5, P≤0.05). These proteins were mainly involved in protein activation, complement activation, immune response, regulation of coagulation and lipoprotein oxidation and other biological processes. The significantly enriched KEGG pathways included the complement and coagulation cascade, phagosome, protein processing in endoplasmic reticulum, antigen processing and presentation, metabolism of glycine, serine and threonine, proteoglycans in cancer, and so on. The interaction analysis of differentially expressed proteins showed that the most significant function modules enriched by differentially expressed proteins were metabolic pathway, complement and coagulation cascades and phagosome. HSP90AA1, HSPA8, APOD, APOM, SERPING1, MASP1, CALR, TUBA1B and TUBB were important nodes in network. There are some differences in serum protein composition between horses and donkeys under physiological conditions. This study provides a foundation for further revealing the differences of physiological characteristics among equine species and effective health protection on them.

Key words: horse, donkey, serum, Label-free, proteomic

中图分类号:

张鏻兮, 韩雨薇, 李政, 廖清超, 汤驰, 邓亮. 基于Label-free技术比较马和驴血清蛋白质组分差异[J]. 畜牧兽医学报, 2021, 52(11): 3099-3107.

ZHANG Linxi, HAN Yuwei, LI Zheng, LIAO Qingchao, TANG Chi, DENG Liang. Comparison of Serum Protein Profiles in Horses and Donkeys Using Label-free Quantitative Proteomics[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(11): 3099-3107.

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