Detection of dimethylarginines in protein hydrolysates by matrix-assisted laser desorption/ionization mass spectrometry |
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Authors: | Hsieh Cheng-Hsilin Tam Ming F |
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Affiliation: | Institute of Molecular Biology, Academia Sinica, NanKang, Taipei, Taiwan, ROC. |
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Abstract: | 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. |
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Keywords: | Arginine methylation Amino acid analysis MALDI-TOF |
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