Immonium ion scanning for the discovery of post-translational modifications and its application to histones.
This study investigates the use of immonium ion scanning for the discovery of methylated and acetylated peptides. Tandem mass spectrometry of modified and unmodified versions of identical peptides revealed ions of 98, 112 and 126 m/ z specifically in association with mono-, dimethylated and acetylated lysine, respectively. Ions of 143 m/ z were seen to be associated with monomethylated arginine, although were not unique to this amino acid. Use of immonium ion scanning with differing collision energies (35, 55, 75, 95, 115 eV) showed that where immonium ions are strong and unique for a modified amino acid, the discovery rate of modified peptides can be improved up to 4-fold over control analyses. The position of an amino acid in a peptide, being terminal or internal, also affected the efficiency of identification of modified peptides. Higher collision energy scanning was required for the most effective identification of peptides with internal modified residues. We conclude that immonium ion scanning, particularly with a range of collision energies, can improve the discovery efficiency of post-translational modifications in peptides.
School of Biotechnology and Biomolecular Sciences and Bioanalytical Mass Spectrometry Facility, University of New South Wales, Sydney, Australia.
Timothy A
TA
Couttas
Mark J
MJ
Raftery
Giulia
G
Bernardini
Marc R
MR
Wilkins
D000107:Acetylation; D000595:Amino Acid Sequence; D002412:Cations; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006657:Histones / Q000032:analysis* / Q000378:metabolism; D007097:Imines / Q000737:chemistry*; D008745:Methylation; D008968:Molecular Conformation; D053758:Nanoparticles; D009842:Oligopeptides / Q000737:chemistry*; D009861:Onium Compounds / Q000737:chemistry*; D011499:Protein Processing, Post-Translational; D053719:Tandem Mass Spectrometry
D002412:Cations; D006657:Histones; D007097:Imines; D009842:Oligopeptides; D009861:Onium Compounds