Double acylation for identification of amino-terminal peptides of proteins isolated by polyacrylamide gel electrophoresis
Article first published online: 14 JUN 2007
Copyright © 2007 John Wiley & Sons, Ltd.
Rapid Communications in Mass Spectrometry
Volume 21, Issue 14, pages 2237–2244, 30 July 2007
How to Cite
Sanchez, A., Ramos, Y., Solano, Y., Gonzalez, L. J., Besada, V., Betancourt, L., Gil, J., Alvarez, F., Rodriguez, M., Perez, L., Pujol, M. and Padron, G. (2007), Double acylation for identification of amino-terminal peptides of proteins isolated by polyacrylamide gel electrophoresis. Rapid Commun. Mass Spectrom., 21: 2237–2244. doi: 10.1002/rcm.3079
- Issue published online: 14 JUN 2007
- Article first published online: 14 JUN 2007
- Manuscript Accepted: 1 MAY 2007
- Manuscript Revised: 27 APR 2007
- Manuscript Received: 23 NOV 2006
We report here a method for the identification of free or blocked N-terminal peptide of in-gel digested isolated proteins. The primary amino groups of the gel-entrapped protein are blocked with normal acetic or succinic anhydride, and the protein is digested with a high-specificity protease. The generated peptides are treated with an equimolar mixture of normal and deuterated acetic anhydride. Upon mass spectrometric analysis internal peptides display a complex isotopic ion distribution while the N-terminal peptide shows a normal isotopic ion distribution. The procedure was applied to the identification of the N-terminus of individual and protein mixtures isolated by sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS-PAGE). Copyright © 2007 John Wiley & Sons, Ltd.