Anhui Key Laboratory of Animal and Poultry Product Safety Engineering, Institute of Animal Science and Veterinary Medicine, Anhui Academy of Agricultural Sciences, Hefei 230031, China.
Anhui Key Laboratory of Animal and Poultry Product Safety Engineering, Institute of Animal Science and Veterinary Medicine, Anhui Academy of Agricultural Sciences, Hefei 230031, China; College of Food Science and Engineering, Qingdao Agricultural University, Qingdao 266109, China.
J Dairy Sci. 2021 Jul;104(7):8265-8275. doi: 10.3168/jds.2020-20097. Epub 2021 Apr 15.
Posttranslational modifications, mostly phosphorylation, are critical for protein structure and function. However, the association between liver phosphoproteins in neonatal calves and colostrum intake is not well understood. In this study, we examined the liver phosphoproteome profile in neonatal calves after receiving colostrum or milk. Liver tissue samples were collected from control calves (CON, n = 3) 2 h after birth and from calves that received colostrum (CG, n = 3) or milk (MG, n = 3) 24 h after birth. Hepatic phosphoprotein expression profiles were analyzed using quantitative proteomics based on the liquid chromatography-tandem mass spectrometry method. In total, 1,587 phosphorylated sites were identified in 1,011 liver proteins. The most abundant phosphorylation site AA was serine (87.5%), followed by threonine (11.9%) and tyrosine (0.5%). Among the 1,011 phosphoproteins, 219, 453, and 26 displayed differential expression in the CG versus MG, CG versus CON, and MG versus CON comparisons, respectively. Differentially expressed phosphoproteins in the CG-MG comparison included 3-phosphoinositide-dependent protein kinase 1, glucose transporter member 4, protein kinase N2, and vinculin, which were mainly involved in the glycogen metabolic process, transport, growth and development, and cell adhesion process, according to Gene Ontology analysis. Pathway analysis indicated their enrichment in the insulin signaling pathway, spliceosome, and adherens junction. The CG-CON comparison identified differentially expressed phosphoproteins and their target genes that were largely involved in the cellular process, macromolecule metabolic process, developmental process, and transport. Pathway analysis indicated their association with endocytosis, mechanistic target of rapamycin, AMP-activated protein kinase, and insulin signaling pathways. These data demonstrate that changes in the phosphoproteins of liver tissues may play an important role in energy metabolism and immune response in the calves that received colostrum. These results provide novel insights into the crucial roles of protein phosphorylation during the early life of newborn calves.
翻译后修饰，主要是磷酸化，对蛋白质的结构和功能至关重要。然而，新生犊牛肝脏磷酸化蛋白质与初乳摄入之间的关联尚未得到充分了解。在本研究中，我们检测了新生犊牛在摄入初乳或牛奶后的肝脏磷酸化蛋白质组概况。从出生后2小时的对照犊牛（CON，n = 3）以及出生后24小时摄入初乳（CG，n = 3）或牛奶（MG，n = 3）的犊牛中采集肝脏组织样本。基于液相色谱 - 串联质谱法，使用定量蛋白质组学分析肝脏磷酸化蛋白质的表达谱。总共在1011种肝脏蛋白质中鉴定出1587个磷酸化位点。最丰富的磷酸化位点氨基酸是丝氨酸（87.5%），其次是苏氨酸（11.9%）和酪氨酸（0.5%）。在1011种磷酸化蛋白质中，分别有219、453和26种在CG与MG、CG与CON、MG与CON的比较中显示出差异表达。根据基因本体分析，CG - MG比较中差异表达的磷酸化蛋白质包括3 - 磷酸肌醇依赖性蛋白激酶1、葡萄糖转运蛋白成员4、蛋白激酶N2和纽蛋白，它们主要参与糖原代谢过程、运输、生长发育和细胞黏附过程。通路分析表明它们在胰岛素信号通路、剪接体和黏着连接中富集。CG - CON比较鉴定出差异表达的磷酸化蛋白质及其靶基因，它们主要参与细胞过程、大分子代谢过程、发育过程和运输。通路分析表明它们与内吞作用、雷帕霉素作用靶点、AMP激活的蛋白激酶和胰岛素信号通路相关。这些数据表明，肝脏组织磷酸化蛋白质的变化可能在摄入初乳的犊牛的能量代谢和免疫反应中发挥重要作用。这些结果为新生犊牛早期生命中蛋白质磷酸化的关键作用提供了新的见解。
