Targeted Proteomic Approaches for Proteome-Wide Characterizations of the AMP-Binding Capacities of Kinases.
Targeted Proteomic Approaches for Proteome-Wide Characterizations of the AMP-Binding Capacities of Kinases.
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DOI：
10.1021/acs.jproteome.2c00225
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发表时间：
2022-08-05
影响因子：
4.4
通讯作者：
Wang, Yinsheng
中科院分区：
文献类型：
--
作者：
Miao, Weili;Yin, Jiekai;Porter, Douglas F.;Jiang, Xiaogang;Khavari, Paul A.;Wang, Yinsheng
激酶在细胞信号传导中起重要作用，已知腺苷一磷酸（AMP）通过AMP活化的蛋白激酶（AMPK）调节细胞能量稳态。在这里，我们探索了新的AMP结合激酶，采用脱硫生物素共轭AMP酰基磷酸探针，有效地富集AMP结合蛋白。结合基于平行反应监测的靶向蛋白质组学方法，我们发现了195种候选AMP结合激酶。我们还丰富了脱硫生物素标记的肽从腺嘌呤核苷酸结合位点的激酶，并分析它们使用LC-MS/MS的多反应监测模式，这导致在鉴定44肽来自43激酶显示可比或更好的结合亲和力对AMP相对于三磷酸腺苷（ATP）。此外，我们的蛋白质组学数据显示，AMP通过直接结合相关激酶，特别是MEK 2和MEK 3，可能参与MAPK通路。总之，我们揭示了大量激酶的AMP结合能力，我们的工作为理解AMP如何作为第二信使调节细胞信号传导奠定了坚实的基础。
Kinases play important roles in cell signaling, and adenosine monophosphate (AMP) is known to modulate cellular energy homeostasis through AMP-activated protein kinase (AMPK). Here, we explored novel AMP-binding kinases by employing a desthiobiotin-conjugated AMP acyl-phosphate probe to enrich efficiently AMP-binding proteins. Together with a parallel-reaction monitoring-based targeted proteomic approach, we uncovered 195 candidate AMP-binding kinases. We also enriched desthiobiotin-labeled peptides from adenine nucleotide-binding sites of kinases and analyzed them using LC-MS/MS in the multiple-reaction monitoring mode, which resulted in the identification of 44 peptides derived from 43 kinases displaying comparable or better binding affinities toward AMP relative to adenosine triphosphate (ATP). Moreover, our proteomic data revealed a potential involvement of AMP in the MAPK pathway through binding directly to the relevant kinases, especially MEK2 and MEK3. Together, we revealed the AMP-binding capacities of a large number of kinases, and our work built a strong foundation for understanding how AMP functions as a second messenger to modulate cell signaling.
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影响因子：
12.8
作者：
Kim J;Yang G;Kim Y;Kim J;Ha J
通讯作者：
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影响因子：
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通讯作者：
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影响因子：
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通讯作者：
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影响因子：
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作者：
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通讯作者：
Wang, Yinsheng