A High-Throughput Targeted Proteomic Approach for Comprehensive Profiling of Methylglyoxal-Induced Perturbations of the Human Kinome
A High-Throughput Targeted Proteomic Approach for Comprehensive Profiling of Methylglyoxal-Induced Perturbations of the Human Kinome
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DOI：
10.1021/acs.analchem.6b02816
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发表时间：
2016-10-04
影响因子：
7.4
通讯作者：
Wang, Yinsheng
中科院分区：
文献类型：
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作者：
Miao, Weili;Xiao, Yongsheng;Wang, Yinsheng
激酶是参与多种细胞信号转导过程的最重要的酶家族之一。现有的研究激酶表达和激活的方法覆盖的范围有限。在这里，我们建立了一种基于多反应监测(MRM)的靶向蛋白质组学方法，该方法提供了前所未有的覆盖(类似于80%)的人类基因组。我们使用这种方法来全面描述在甲基乙二醛治疗后HEK293T人类胚胎肾细胞的全球基因组的变化，甲乙二醛是一种糖酵解副产物，在糖尿病患者的血液和组织中存在高水平，被认为是导致糖尿病并发症的原因之一。我们的结果导致了328个独特的激酶的量化。特别是，我们发现甲基乙二醛处理引起了MAPK通路中一些激酶的表达改变，并减少了几种受体酪氨酸激酶的表达，包括表皮生长因子受体(EGFR)、胰岛素生长因子2受体(IGF2R)、成纤维细胞生长因子受体(FGFR)等。此外，我们还证明了EGFR的表达减少是通过一种与IGF2R和FGFR1不同的机制发生的。总之，我们的靶向动态组分析方法为探索被细胞外刺激改变的激酶介导的信号通路提供了强大的资源；本研究的结果提示了糖尿病并发症发生的新机制。
Kinases are one of the most important families of enzymes that are involved in numerous cell signaling processes. Existing methods for studying kinase expression and activation have limited kinome coverage. Herein we established a multiple-reaction monitoring (MRM)-based targeted proteomic method that provided an unprecedented coverage (similar to 80%) of the human kinome. We employed this method for profiling comprehensively the alterations of the global kinome of HEK293T human embryonic kidney cells upon treatment with methylglyoxal, a glycolysis by product that is present at elevated levels in blood and tissues of diabetic patients and is thought to contribute to diabetic complications. Our results led to the quantification of 328 unique kinases. In particular, we found that methylglyoxal treatment gave rise to altered expression of a number of kinases in the MAPK pathway and diminished expression of several receptor tyrosine kinases, including epidermal growth factor receptor (EGFR), insulin growth factor 2 receptor (IGF2R), fibroblast growth factor receptor (FGFR), etc. Furthermore, we demonstrated that the diminished expression of EGFR occurred through a mechanism that is distinct from the reduced expression of IGF2R and FGFR1. Together, our targeted kinome profiling method offers a powerful resource for exploring kinase-mediated signaling pathways that are altered by extracellular stimuli; and the results from the, present study suggest new mechanisms underlying the development of diabetic complications.