Correct identification of oxidized histidine residues using electron-transfer dissociation.
Oxidative modification to the side chain of histidine can noticeably change the collision-induced dissociation (CID) pathways of peptides containing this oxidized residue. In cases where an oxidized peptide consists two or more isomers differing only in the site of modification, oxidation to histidine usually causes the other oxidized sites to be mis-assigned in CID spectra. These spectral misassignments can sometimes be avoided by using multiple stages of MS/MS (MS(n)) or via specially optimized liquid chromatographic separation conditions. In this manuscript, we demonstrate that these misassignments can be more readily and easily avoided by using electron-transfer dissociation (ETD) to dissociate the oxidized peptides. Furthermore, we find that the relative insensitivity of ETD to side-chain chemistry allows the extent of oxidative modification to be determined readily for peptide isomers having more than one site of oxidation. The current results along with previous studies of oxidized peptides suggest that ETD is probably a better technique than CID for obtaining correct sequence and modification information for oxidized peptides.
Srikanth
Rapole
R
Department of Chemistry, University of Massachusetts, Amherst, Massachusetts 01003, USA.
Wilson
Jonathan
J
Vachet
Richard W
RW
D004583:Electrons; D006639:Histidine / Q000737:chemistry* / Q000378:metabolism; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry; D020539:Sequence Analysis, Protein; D053719:Tandem Mass Spectrometry / Q000379:methods*
D010455:Peptides; D011506:Proteins; D006639:Histidine