A mass spectrometry based method for distinguishing between symmetrically and asymmetrically dimethylated arginine residues.

The arginine methylation of proteins is involved in several important cellular activities, most notably transcriptional control. Arginine dimethylation can take two distinct forms, symmetric and asymmetric, catalyzed by different classes of enzymes. To establish a method for the mass spectrometric identification and characterization of this post-translational modification, we analyzed synthetic peptides with symmetrically or asymmetrically methylated arginine residues by electrospray ionization tandem mass spectrometry. We observed abundant characteristic ions at M+nH-31 and M+nH-70 in spectra of symmetrically methylated peptides and at M+nH-45 in spectra of asymmetrically methylated peptides. We speculate these ions arise from neutral loss of monomethylamine, dimethylcarbodiimide, and dimethylamine, respectively. These characteristic ions allowed the rapid identification of a symmetrically arginine-dimethylated peptide from myelin basic protein and a symmetrically arginine-dimethylated peptide from SmD3 co-immunoprecipitated with the methyltransferase-associated protein pICln, suggesting that this method may provide a rapid means to screen for and characterize dimethylarginine sites.