人类胚胎干细胞的磷酸化蛋白质组学分析
Phosphoproteomic analysis of human embryonic stem cells.
作者信息
Brill Laurence M, Xiong Wen, Lee Ki-Bum, Ficarro Scott B, Crain Andrew, Xu Yue, Terskikh Alexey, Snyder Evan Y, Ding Sheng
机构信息
Genomics Institute of the Novartis Research Foundation, San Diego, CA 92109, USA.
出版信息
Cell Stem Cell. 2009 Aug 7;5(2):204-13. doi: 10.1016/j.stem.2009.06.002.
Protein phosphorylation, while critical to cellular behavior, has been undercharacterized in pluripotent cells. Therefore, we performed phosphoproteomic analyses of human embryonic stem cells (hESCs) and their differentiated derivatives. A total of 2546 phosphorylation sites were identified on 1602 phosphoproteins; 389 proteins contained more phosphorylation site identifications in undifferentiated hESCs, whereas 540 contained more such identifications in differentiated derivatives. Phosphoproteins in receptor tyrosine kinase (RTK) signaling pathways were numerous in undifferentiated hESCs. Cellular assays corroborated this observation by showing that multiple RTKs cooperatively supported undifferentiated hESCs. In addition to bFGF, EGFR, VEGFR, and PDGFR activation was critical to the undifferentiated state of hESCs. PDGF-AA complemented a subthreshold bFGF concentration to maintain undifferentiated hESCs. Also consistent with phosphoproteomics, JNK activity participated in maintenance of undifferentiated hESCs. These results support the utility of phosphoproteomic data, provide guidance for investigating protein function in hESCs, and complement transcriptomics/epigenetics for broadening our understanding of hESC fate determination.
蛋白质磷酸化虽然对细胞行为至关重要，但在多能细胞中的研究尚不充分。因此，我们对人类胚胎干细胞（hESC）及其分化衍生物进行了磷酸化蛋白质组分析。在1602种磷酸化蛋白质上共鉴定出2546个磷酸化位点；389种蛋白质在未分化的hESC中含有更多的磷酸化位点鉴定，而540种蛋白质在分化衍生物中含有更多此类鉴定。受体酪氨酸激酶（RTK）信号通路中的磷酸化蛋白质在未分化的hESC中数量众多。细胞实验通过表明多种RTK协同支持未分化的hESC证实了这一观察结果。除了bFGF外，EGFR、VEGFR和PDGFR的激活对hESC的未分化状态至关重要。PDGF-AA补充亚阈值bFGF浓度以维持未分化的hESC。同样与磷酸化蛋白质组学一致，JNK活性参与维持未分化的hESC。这些结果支持了磷酸化蛋白质组数据的实用性，为研究hESC中的蛋白质功能提供了指导，并补充了转录组学/表观遗传学，以拓宽我们对hESC命运决定的理解。
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