A Selenium-Based Click AdoMet Analogue for Versatile Substrate Labeling with Wild-Type Protein Methyltransferases
A Selenium-Based Click AdoMet Analogue for Versatile Substrate Labeling with Wild-Type Protein Methyltransferases
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DOI：
10.1002/cbic.201100781
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发表时间：
2012-05-29
期刊：
影响因子：
3.2
通讯作者：
Weinhold, Elmar
中科院分区：
文献类型：
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作者：
Willnow, Sophie;Martin, Michael;Weinhold, Elmar
蛋白质甲基化是由s -腺苷- l-蛋氨酸依赖的蛋白质甲基转移酶（MTases）催化的，这种翻译后修饰具有多种细胞功能。一些mtase似乎表现出广泛的底物特异性，因此需要综合的靶谱分析方法。在这里，我们报道了一种新的AdoMet类似物的合成，用于通过铜催化叠氮烷环加成（CuAAC）转移小丙炔基团并标记修饰的蛋白质。硒对硫的替代极大地增强了前甘酸辅因子的稳定性，结合硒离子中心更好的活化，导致更高的酶反应活性。广泛的作用于赖氨酸，精氨酸和谷氨酰胺残基的野生型蛋白mtase接受这种辅助因子，并且修饰的底物可以通过CuAAC点击化学有效地标记。
Protein methylation is catalyzed by S-adenosyl-L-methionine-dependent protein methyltransferases (MTases), and this posttranslational modification serves diverse cellular functions. Some MTases seem to exhibit broad substrate specificities and comprehensive methods for target profiling are needed. Here we report the synthesis of a new AdoMet analogue for enzymatic transfer of a small propargyl group and labeling of modified proteins through copper-catalyzed azidealkyne cycloaddition (CuAAC). Replacement of sulfur by selenium strongly enhanced the stability of the progargylic cofactor, leading, in combination with better activation by the selenonium center, to higher enzymatic reactivity. A broad spectrum of wild-type protein MTases acting on lysine, arginine, and glutamine residues accept this cofactor and modified substrates can be efficiently labeled by CuAAC click chemistry.