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Double acylation for identification of amino‐terminal peptides of proteins isolated by polyacrylamide gel electrophoresis
Author(s) -
Sanchez Aniel,
Ramos Yassel,
Solano Yanni,
Gonzalez Luis Javier,
Besada Vladimir,
Betancourt Lazaro,
Gil Jeovanis,
Alvarez Felix,
Rodriguez Meilyn,
Perez Lincidio,
Pujol Merardo,
Padron Gabriel
Publication year - 2007
Publication title -
rapid communications in mass spectrometry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.528
H-Index - 136
eISSN - 1097-0231
pISSN - 0951-4198
DOI - 10.1002/rcm.3079
Subject(s) - chemistry , chromatography , acylation , acetic anhydride , peptide , succinic anhydride , polyacrylamide gel electrophoresis , gel electrophoresis , biochemistry , organic chemistry , enzyme , catalysis
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.