An extensive survey of tyrosine phosphorylation revealing new sites in human mammary epithelial cells.
An extensive survey of tyrosine phosphorylation revealing new sites in human mammary epithelial cells.
复制标题
DOI：
10.1021/pr900044c
复制
发表时间：
2009-08
影响因子：
4.4
通讯作者：
Qian WJ
中科院分区：
文献类型：
--
作者：
Heibeck TH;Ding SJ;Opresko LK;Zhao R;Schepmoes AA;Yang F;Tolmachev AV;Monroe ME;Camp DG 2nd;Smith RD;Wiley HS;Qian WJ
蛋白酪氨酸磷酸化是细胞信号传导的一个重要调控机制。在这里，我们通过应用抗磷酸酪氨酸肽免疫亲和纯化和高灵敏度毛细管液相色谱串联质谱，对正常来源的人乳腺上皮细胞系中的酪氨酸磷酸化位点进行了广泛的调查。在基础条件和表皮生长因子（EGF）的急性刺激分析后，HMEC确定了285种蛋白质的481个酪氨酸磷酸化位点（由716个独特的肽覆盖）。估计的错误发现率为1.0%，这是通过对一个混乱的数据库进行搜索确定的。将这些数据与现有文献进行比较，发现与先前报道的位点有显著的一致性。然而，我们观察到281个位点以前未在HMEC培养中报道，其中29个位点未在任何人类细胞或组织系统中报道。分析表明，大部分高度磷酸化的蛋白丰度相对较低。我们还观察到，同一蛋白中不同位点的磷酸化化学计量差异很大，这就提出了pTyr高度磷酸化位点可能具有更重要的功能作用。通过映射到主要的信号网络，如EGF受体和胰岛素生长因子-1受体信号通路，揭示了许多已知的参与这些通路的蛋白质是酪氨酸磷酸化的，这为未来的假设驱动和靶向定量研究提供了有趣的靶点，涉及HMEC或其他人体系统中的酪氨酸磷酸化。
Protein tyrosine phosphorylation represents a central regulatory mechanism in cell signaling. Here we present an extensive survey of tyrosine phosphorylation sites in a normal-derived human mammary epithelial cell line by applying anti-phosphotyrosine peptide immunoaffinity purification coupled with high sensitivity capillary liquid chromatography tandem mass spectrometry. A total of 481 tyrosine phosphorylation sites (covered by 716 unique peptides) from 285 proteins were confidently identified in HMEC following the analysis of both the basal condition and acute stimulation with epidermal growth factor (EGF). The estimated false discovery rate was 1.0% as determined by searching against a scrambled database. Comparison of these data with existing literature showed significant agreement for previously reported sites. However, we observed 281 sites that were not previously reported for HMEC cultures and 29 of which have not been reported for any human cell or tissue system. The analysis showed that the majority of highly phosphorylated proteins were relatively low-abundance. Large differences in phosphorylation stoichiometry for sites within the same protein were also observed, raising the possibility of more important functional roles for such highly phosphorylated pTyr sites. By mapping to major signaling networks, such as the EGF receptor and insulin growth factor-1 receptor signaling pathways, many known proteins involved in these pathways were revealed to be tyrosine phosphorylated, which provides interesting targets for future hypothesis-driven and targeted quantitative studies involving tyrosine phosphorylation in HMEC or other human systems.
登录
查看更多内容
影响因子：
4.8
作者：
Lee, In Hye;Lim, Hee Jung;Bae, Yun Soo
通讯作者：
Bae, Yun Soo
影响因子：
46.9
作者：
Blagoev, B;Ong, SE;Mann, M
通讯作者：
Mann, M
影响因子：
4.8
作者：
Katoh, Y;Shiba, Y;Nakayama, K
通讯作者：
Nakayama, K
影响因子：
5.3
作者：
Law, SF;O'Neill, GM;Golemis, EA
通讯作者：
Golemis, EA
影响因子：
46.9
作者：
Ficarro, SB;McCleland, ML;White, FM
通讯作者：
White, FM