Signaling initiated by overexpression of the fibroblast growth factor receptor-1 investigated by mass spectrometry
Signaling initiated by overexpression of the fibroblast growth factor receptor-1 investigated by mass spectrometry
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DOI：
10.1074/mcp.m200075-mcp200
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发表时间：
2003-01-01
影响因子：
7
通讯作者：
Mann, M
中科院分区：
文献类型：
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作者：
Hinsby, AM;Olsen, JV;Mann, M
成纤维细胞生长因子受体-1（FGFR-1）（一种原型受体酪氨酸激酶）的过表达是几种人类肿瘤的特征。在人293细胞中，FGFR-1的过表达导致受体的组成性激活，伴随着含磷酸酪氨酸蛋白的细胞水平的持续高增加。在这里，我们使用质谱法研究FGFR-1过表达诱导的酪氨酸磷酸化蛋白。FGFR-1信号传导的几个众所周知的组分被鉴定为沿着两个新的候选物：NS-1相关蛋白-1和Myb 1样蛋白的靶标。随后，我们应用质谱前体离子扫描，以确定22个酪氨酸磷酸化位点分布在6个底物蛋白的FGFR-1或下游酪氨酸激酶。新的体内酪氨酸磷酸化位点被发现在FGFR-1，磷脂酶C γ，p90核糖体S6激酶，corneum，NS-1相关蛋白-1作为持续的FGFR-1信号的结果，我们提出这些作为功能链接到下游分子和细胞过程。
Overexpression of the fibroblast growth factor receptor-1 (FGFR-1), a prototypic receptor tyrosine kinase, is a feature of several human tumors. In human 293 cells overexpression of the FGFR-1 leads to constitutive activation of the receptor with concomitant sustained high increase in the cellular level of phosphotyrosine-containing proteins. Here we use mass spectrometry to study the tyrosine-phosphorylated proteins induced by overexpression of the FGFR-1. Several well known components of FGFR-1 signaling were identified along with two novel candidates: NS-1-associated protein-1 and target of Myb 1-like protein. We subsequently applied mass spectrometry precursor ion scanning to identify 22 tyrosine phosphorylation sites distributed on six substrate proteins of the FGFR-1 or downstream tyrosine kinases. Novel in vivo tyrosine phosphorylation sites were found in the FGFR-1, phospholipase Cgamma, p90 ribosomal S6 kinase, cortactin, and NS-1-associated protein-1 as a result of sustained FGFR-1 signaling, and we propose these as functional links to downstream molecular and cellular processes.