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

• Published in: Amino acids Vol. 47; no. 12; pp. 2561 - 2571
• Main Authors: Lakowski, Ted M, Pak, Magnolia L, Szeitz, András, Thomas, Dylan, Vhuiyan, Mynol I, Clement, Bernd, Frankel, Adam
• Format: Journal Article
• Language: English
• Published: Vienna Springer Vienna 01.12.2015; Springer Nature B.V
• Subjects: Amino acids; Analytical Chemistry; arginine; Arginine - analogs & derivatives; Arginine - chemistry; Biochemical Engineering; Biochemistry; Biomedical and Life Sciences; Deletion; Gene Deletion; heat; Heat shock; Heat-Shock Response; Hot Temperature; ions; Life Sciences; Mass Spectrometry; Methylation; methyltransferases; Monitoring; Neurobiology; omega-N-Methylarginine - chemistry; Original Article; Protein-Arginine N-Methyltransferases - chemistry; Proteins; Proteomics; Saccharomyces cerevisiae - chemistry; Saccharomyces cerevisiae Proteins - chemistry; Spectra; Yeast; yeasts; sDMA; Yeast; Log phase; ADMA; Protein methylation; MRM; Mass spectrometry; Delta-monomethylarginine; Heat shock; Stationary phase; MRM3
• ISSN: 0939-4451; 1438-2199
• DOI: 10.1007/s00726-015-2047-5

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 Nη1, Nη2-dimethylarginine (sDMA) is present on yeast proteins, and make a detailed structural determination of the fragment ions from the spectra. Nη-monomethylarginine (ηMMA), Nδ-monomethylarginine (δMMA), Nη1, Nη1-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 ηMMA and δMMA 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, ηMMA, and δMMA on yeast proteins.