Peptide and protein sequence analysis by electron transfer dissociation mass spectrometry.
Peptide sequence analysis using a combination of gas-phase ion/ion chemistry and tandem mass spectrometry (MS/MS) is demonstrated. Singly charged anthracene anions transfer an electron to multiply protonated peptides in a radio frequency quadrupole linear ion trap (QLT) and induce fragmentation of the peptide backbone along pathways that are analogous to those observed in electron capture dissociation. Modifications to the QLT that enable this ion/ion chemistry are presented, and automated acquisition of high-quality, single-scan electron transfer dissociation MS/MS spectra of phosphopeptides separated by nanoflow HPLC is described.
Syka
John E P
JE
Engineering Physics Program, University of Virginia, Charlottesville, VA 22901, USA.
Coon
Joshua J
JJ
Schroeder
Melanie J
MJ
Shabanowitz
Jeffrey
J
Hunt
Donald F
DF
D000595:Amino Acid Sequence; D000838:Anions / Q000737:chemistry; D000873:Anthracenes / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D004579:Electron Transport; D004583:Electrons; D013058:Mass Spectrometry / Q000379:methods*; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification; D010750:Phosphoproteins / Q000737:chemistry* / Q000302:isolation & purification; D020539:Sequence Analysis, Protein / Q000379:methods*
D000838:Anions; D000873:Anthracenes; D010455:Peptides; D010750:Phosphoproteins; C034020:anthracene