Detection of dimethylarginines in protein hydrolysates by matrix-assisted laser desorption/ionization mass spectrometry.
We report a method to detect the presence of dimethylarginines on proteins. Peptides with dimethylarginines were hydrolyzed in acid. The hydrolysates were subjected to matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometric analysis using a mixture of alpha-cyano-4-hydroxycinnamic acid and nitrocellulose as matrix. Both asymmetric omega-N(G),N(G)-dimethylarginine and symmetric omega-N(G),N(G')-dimethylarginine give a clear signal at m/z 203. Recombinant Sbp1p modified by Hmt1p in vivo were isolated by affinity chromatography followed by electrophoresis on a polyacrylamide gel and subjected to acid hydrolysis. MALDI-TOF analysis of the acid hydrolysates confirmed the presence of dimethylarginines. The detection limit of the method is estimated at approximately 1pmol of protein.
Hsieh
Cheng-Hsilin
CH
Institute of Molecular Biology, Academia Sinica, NanKang, Taipei, Taiwan, ROC.
Tam
Ming F
MF
D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry; D001798:Blood Proteins / Q000737:chemistry; D003101:Collodion / Q000737:chemistry; D003373:Coumaric Acids / Q000737:chemistry; D029941:Fish Proteins / Q000737:chemistry; D011492:Protein Hydrolysates / Q000737:chemistry; D011484:Protein-Arginine N-Methyltransferases / Q000378:metabolism; D012097:Repressor Proteins / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D001798:Blood Proteins; D003373:Coumaric Acids; D029941:Fish Proteins; D011492:Protein Hydrolysates; D012097:Repressor Proteins; C088488:SBP1 protein, Parablax neblifer; D029701:Saccharomyces cerevisiae Proteins; C487735:dimethylarginine; C007175:alpha-cyano-4-hydroxycinnamate; C018524:N,N-dimethylarginine; D003101:Collodion; D001120:Arginine; C475182:HMT1 protein, S cerevisiae; D011484:Protein-Arginine N-Methyltransferases