Detection of dimethylarginines in protein hydrolysates by matrix-assisted laser desorption/ionization mass spectrometry.
Anal Biochem
; 350(1): 151-5, 2006 Mar 01.
Article
em En
| MEDLINE
| ID: mdl-16403427
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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Base de dados:
MEDLINE
Assunto principal:
Arginina
/
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
Idioma:
En
Ano de publicação:
2006
Tipo de documento:
Article