Phosphoproteomic analysis of human embryonic stem cells.
Phosphoproteomic analysis of human embryonic stem cells.
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DOI：
10.1016/j.stem.2009.06.002
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发表时间：
2009-08-07
期刊：
影响因子：
23.9
通讯作者：
Ding S
中科院分区：
文献类型：
--
作者：
Brill LM;Xiong W;Lee KB;Ficarro SB;Crain A;Xu Y;Terskikh A;Snyder EY;Ding S
蛋白质磷酸化虽然对细胞行为至关重要，但在多能细胞中尚未得到充分表征。因此，我们对人胚胎干细胞（hESC）及其分化衍生物进行了磷酸化蛋白质组学分析。在 1602 个磷蛋白上鉴定出 2546 个磷酸化位点； 389 个蛋白质在未分化 hESC 中包含更多磷酸化位点识别，而 540 个蛋白质在分化衍生物中包含更多此类识别。未分化 hESC 中受体酪氨酸激酶 (RTK) 信号通路中的磷蛋白数量众多。细胞检测证实了这一观察结果，表明多个 RTK 协同支持未分化的 hESC。除了 bFGF 之外，EGFR、VEGFR 和 PDGFR 的激活对于 hESC 的未分化状态也至关重要。 PDGF-AA 补充阈值以下的 bFGF 浓度以维持未分化的 hESC。同样与磷酸化蛋白质组学一致的是，JNK 活性参与了未分化 hESC 的维持。这些结果支持磷酸化蛋白质组学数据的实用性，为研究 hESC 中已知和新型蛋白质提供指导，并补充转录组学/表观遗传学以扩大我们对 hESC 命运决定的理解。
Protein phosphorylation, while critical to cellular behavior, has been under-characterized in pluripotent cells. Therefore, we performed phosphoproteomic analyses of human embryonic stem cells (hESCs) and their differentiated derivatives. 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 sub-threshold 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 known and novel proteins in hESCs, and complement transcriptomics/epigenetics for broadening our understanding of hESC fate determination.
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DOI：
10.1073/pnas.251194298
发表时间：
2001-11-20
影响因子：
11.1
作者：
Bennett, BL;Sasaki, DT;Anderson, DW
通讯作者：
Anderson, DW
影响因子：
64.5
作者：
Lee, TI;Jenner, RG;Young, RA
通讯作者：
Young, RA
影响因子：
7.4
作者：
Liu, HB;Sadygov, RG;Yates, JR
通讯作者：
Yates, JR
影响因子：
56.9
作者：
Cha, TL;Zhou, BHP;Hung, MC
通讯作者：
Hung, MC
影响因子：
48
作者：
Bodenmiller, Bernd;Mueller, Lukas N.;Aebersold, Ruedi
通讯作者：
Aebersold, Ruedi