Quantitative Analysis and Discovery of Lysine and Arginine Modifications

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• 作者：James J. GalliganPhilip J. Kingsley ; Orrette R. Wauchope ; Michelle M. Mitchener ; Jeannie M. Camarillo ; James A. WepyPeter S. Harris ; Kristofer S. Fritz ; Lawrence J. Marnett
• 刊名：Analytical Chemistry
• 出版年：2017
• 出版时间：January 17, 2017
• 年：2017
• 卷：89
• 期：2
• 页码：1299-1306
• 全文大小：451K
• ISSN：1520-6882

文摘

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.