An Azo Coupling-Based Chemoproteomic Approach to Systematically Profile the Tyrosine Reactivity in the Human Proteome.
An Azo Coupling-Based Chemoproteomic Approach to Systematically Profile the Tyrosine Reactivity in the Human Proteome.
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DOI：
10.1021/acs.analchem.1c01935
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发表时间：
2021-07-27
影响因子：
7.4
通讯作者：
Wu R
中科院分区：
文献类型：
--
作者：
Sun F;Suttapitugsakul S;Wu R
蛋白质的酪氨酸残基参与广泛的活性，包括酶催化、蛋白质-蛋白质相互作用和蛋白质-配体结合。然而，大规模的酪氨酸残基的功能注释仍然非常具有挑战性。在这里，我们报告了一种新的方法，结合偶氮偶联，生物正交化学，和多路复用蛋白质组学，在全球范围内调查酪氨酸在人类蛋白质组的反应性。基于芳基重氮盐与酪氨酸残基的偶氮偶联反应，对两种不同的探针进行了评价，并选择性能最好的探针用于进一步研究人类蛋白质组中的酪氨酸残基。然后，标记的含酪氨酸的肽选择性富集使用生物正交化学，切割后，肽上的小标签完全适合通过MS进行位点特异性分析。与多路复用蛋白质组学耦合，我们量化了MCF 7细胞中超过5000个酪氨酸位点，这些量化的位点显示出广泛的反应性。具有高反应活性的酪氨酸残基存在于功能和结构多样的蛋白质上，包括具有催化活性和结合性质的蛋白质。这种方法可以广泛应用于推进我们对蛋白质功能的理解，并促进开发共价药物来调节蛋白质活性。
The tyrosine residue of proteins participates in a wide range of activities including enzymatic catalysis, protein–protein interaction, and protein–ligand binding. However, the functional annotation of the tyrosine residues on a large scale is still very challenging. Here, we report a novel method integrating azo coupling, bioorthogonal chemistry, and multiplexed proteomics to globally investigate the tyrosine reactivity in the human proteome. Based on the azo-coupling reaction between aryl diazonium salt and the tyrosine residue, two different probes were evaluated, and the probe with the best performance was employed to further study the tyrosine residues in the human proteome. Then, tagged tyrosine-containing peptides were selectively enriched using bioorthogonal chemistry, and after the cleavage, a small tag on the peptides perfectly fits for site-specific analysis by MS. Coupling with multiplexed proteomics, we quantified over 5000 tyrosine sites in MCF7 cells, and these quantified sites displayed a wide range of reactivity. The tyrosine residues with high reactivity were found on functionally and structurally diverse proteins, including those with the catalytic activity and binding property. This method can be extensively applied to advance our understanding of protein functions and facilitate the development of covalent drugs to regulate protein activity.
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影响因子：
6.6
作者：
Huang M;Wang Y
通讯作者：
Wang Y
影响因子：
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作者：
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通讯作者：
Wu R
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通讯作者：
Francis, MB
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通讯作者：
Sidhu, Sachdev S.
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通讯作者：
Chatterjee A