An NMR-Based Approach for Global Arginine Methylation Analysis.

Protein arginine methylation (ArgMet) plays a crucial role in the regulation of cellular processes, including transcription, RNA processing, signal transduction, and DNA damage response. However, the mechanisms linking protein ArgMet dynamics to (patho)physiology remain unclear due to the lack of global analysis methods. In this chapter, we present a robust protocol for the quantification of global protein ArgMet, including asymmetric dimethylarginine, symmetric dimethylarginine, and monomethylarginine. After isolation of proteins from biological fluids, tissues, or cell lysates, a straightforward method for homogenization, precipitation, and hydrolysis of proteins is outlined. As the hydrolysates contain a high variability of components, nuclear magnetic resonance (NMR)-based detection was used, providing a robust tool to study arginine methylome with high specificity.