Arginine methylation in yeast proteins during stationary-phase growth and heat shock

作者：Lakowski Ted M^*; Pak Magnolia L; Szeitz Andars; Thomas Dylan; Vhuiyan Mynol I; Clement Bernd; Frankel Adam

来源：Amino Acids, 2015, 47(12): 2561-2571.

DOI：10.1007/s00726-015-2047-5

• ADMA
• sDMA
• Yeast
• Mass spectrometry
• Protein methylation
• Delta-monomethylarginine
• Heat shock
• Stationary phase
• Log phase
• MRM3

Arginine methyltransferases (RMTs) catalyze the methylation of arginine residues on proteins. We examined the effects of log-phase growth, stationary-phase growth, and heat shock on the formation of methylarginines on yeast proteins to determine if the conditions favor a particular type of methylation. Utilizing linear ion trap mass spectrometry, we identify methylarginines in wild-type and RMT deletion yeast strains using secondary product ion scans (MS3), and quantify the methylarginines using multiple reaction monitoring (MRM). Employing MS3 and isotopic incorporation, we demonstrate for the first time that N eta 1, N eta 2-dimethylarginine (sDMA) is present on yeast proteins, and make a detailed structural determination of the fragment ions from the spectra. N eta-monomethylarginine (eta MMA), N delta-monomethylarginine (delta MMA), N eta 1, N eta 1-dimethylarginine (aDMA), and sDMA were detected in RMT deletion yeast using MS3 and MRM with and without isotopic incorporation, suggesting that additional RMT enzymes remain to be discovered in yeast. The concentrations of eta MMA and delta MMA decreased by half during heat shock and stationary phase compared to log-phase growth of wild-type yeast, whereas sDMA increased by as much as sevenfold and aDMA decreased by 11-fold. Therefore, upon entering stressful conditions like heat shock or stationary-phase growth, there is a net increase in sDMA and decreases in aDMA, eta MMA, and delta MMA on yeast proteins.

• 出版日期2015-12