title: Double acylation for identification of amino-terminal peptides of proteins isolated by polyacrylamide gel electrophoresis.
abstract: 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).
authors.affiliation: Department of Proteomics, Center for Genetic Engineering and Biotechnology, Havana, Cuba.
authors.forename: Aniel
authors.initials: A
authors.lastname: Sanchez
authors.forename: Yassel
authors.initials: Y
authors.lastname: Ramos
authors.forename: Yanni
authors.initials: Y
authors.lastname: Solano
authors.forename: Luis Javier
authors.initials: LJ
authors.lastname: Gonzalez
authors.forename: Vladimir
authors.initials: V
authors.lastname: Besada
authors.forename: Lazaro
authors.initials: L
authors.lastname: Betancourt
authors.forename: Jeovanis
authors.initials: J
authors.lastname: Gil
authors.forename: Felix
authors.initials: F
authors.lastname: Alvarez
authors.forename: Meilyn
authors.initials: M
authors.lastname: Rodriguez
authors.forename: Lincidio
authors.initials: L
authors.lastname: Perez
authors.forename: Merardo
authors.initials: M
authors.lastname: Pujol
authors.forename: Gabriel
authors.initials: G
authors.lastname: Padron
mesh_terms: D000215:Acylation; D000596:Amino Acids / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods; D010449:Peptide Mapping / Q000379:methods*; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D013194:Staining and Labeling / Q000379:methods
chemical_list: D000596:Amino Acids; D010455:Peptides; D011506:Proteins