Dynamic Phosphoproteome Profiling of Zebrafish Embryonic Fibroblasts during Cold Acclimation
Dynamic Phosphoproteome Profiling of Zebrafish Embryonic Fibroblasts during Cold Acclimation
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冷驯化过程中斑马鱼胚胎成纤维细胞的动态磷酸化蛋白质组分析
DOI：
10.1002/pmic.201900257
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发表时间：
2020
期刊：
影响因子：
3.4
通讯作者：
Zongbin Cui
中科院分区：
文献类型：
--
作者：
Junjun Yan;Yong Long;Tong Zhou;Jing Ren;Qing Li;Guili Song;Zongbin Cui
温度几乎影响鱼类生活的各个方面。为了应对低温，鱼类进化出了适应寒冷生存的能力。然而，鱼类冷驯化背后的细胞内信号传导事件仍然很大程度上未知。在这里，监测斑马鱼胚胎成纤维细胞（ZF4）冷驯化的形成，并通过大规模定量磷酸化蛋白质组学方法研究该过程中的磷酸化事件。总共在 4066 个蛋白质上鉴定出 11,474 个磷酸化位点，并在 2519 个蛋白质上定量了 5772 个磷酸化位点。丝氨酸、苏氨酸和酪氨酸 (Ser/Thr/Tyr) 磷酸化分别占总磷酸位点的 85.5%、13.3% 和 1.2%。在所有磷酸位点中，510 个蛋白质上的 702 个磷酸位点在 ZF4 细胞冷驯化过程中显示出差异调节。这些磷酸位点根据其在冷暴露过程中的动态变化分为六簇。激酶底物预测表明，激酶组中的丝裂原激活蛋白激酶 (MAPK) 主要负责这些磷酸位点的磷酸化。差异调节的磷蛋白在功能上与各种细胞过程相关，例如肌动蛋白细胞骨架和 MAPK 信号通路的调节。这些数据丰富了斑马鱼蛋白质磷酸化位点的数据库，并为阐明鱼类冷驯化过程中的细胞内信号网络提供了关键线索。
Temperature affects almost all aspects of the fish life. To cope with low temperature, fish have evolved the ability of cold acclimation for survival. However, intracellular signaling events underlying cold acclimation in fish remain largely unknown. Here, the formation of cold acclimation in zebrafish embryonic fibroblasts (ZF4) is monitored and the phosphorylation events during the process are investigated through a large‐scale quantitative phosphoproteomic approach. In total, 11 474 phosphorylation sites are identified on 4066 proteins and quantified 5772 phosphosites on 2519 proteins. Serine, threonine, and tyrosine (Ser/Thr/Tyr) phosphorylation accounted for 85.5%, 13.3%, and 1.2% of total phosphosites, respectively. Among all phosphosites, 702 phosphosites on 510 proteins show differential regulation during cold acclimation of ZF4 cells. These phosphosites are divided into six clusters according to their dynamic changes during cold exposure. Kinase–substrate prediction reveals that mitogen‐activated protein kinase (MAPK) among the kinase groups is predominantly responsible for phosphorylation of these phosphosites. The differentially regulated phosphoproteins are functionally associated with various cellular processes such as regulation of actin cytoskeleton and MAPK signaling pathway. These data enrich the database of protein phosphorylation sites in zebrafish and provide key clues for the elucidation of intracellular signaling networks during cold acclimation of fish.