Quantitative Analysis and Discovery of Lysine and Arginine Modifications.
Quantitative Analysis and Discovery of Lysine and Arginine Modifications.
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DOI：
10.1021/acs.analchem.6b04105
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发表时间：
2017-01-17
影响因子：
7.4
通讯作者：
Marnett LJ
中科院分区：
文献类型：
--
作者：
Galligan JJ;Kingsley PJ;Wauchope OR;Mitchener MM;Camarillo JM;Wepy JA;Harris PS;Fritz KS;Marnett LJ
翻译后修饰（PTM）影响蛋白质的功能，定位和稳定性，但很少有人知道这些修饰的比例。在这里，我们描述了一种新的方法来定量和评估的相对化学计量的赖氨酸和精氨酸的修改（QuARKMod）在复杂的生物设置。我们证明了该平台在监测重组蛋白修饰肽底物，PTMs的个别组蛋白，这些PTMs的相对丰度作为亚细胞位置的函数的多功能性。最后，我们描述了一个产品离子扫描技术，提供了潜在的发现意想不到的和可能新颖的赖氨酸和精氨酸的修改。总之，这种方法在复杂的生物系统中产生蛋白质PTM的准确定量和发现，而不需要高质量精度的仪器。
Post-translational modifications (PTMs) affect protein function, localization, and stability, yet very little is known about the ratios of these modifications. Here, we describe a novel method to quantitate and assess the relative stoichiometry of Lys and Arg modifications (QuARKMod) in complex biological settings. We demonstrate the versatility of this platform in monitoring recombinant protein modification of peptide substrates, PTMs of individual histones, and the relative abundance of these PTMs as a function of subcellular location. Lastly, we describe a product ion scanning technique that offers the potential to discover unexpected and possibly novel Lys and Arg modifications. In summary, this approach yields accurate quantitation and discovery of protein PTMs in complex biological systems without the requirement of high mass accuracy instrumentation.