Rapidly alternating transmission mode electron-transfer dissociation and collisional activation for the characterization of polypeptide ions.
Cation transmission/electron-transfer reagent anion storage mode electron-transfer ion/ion reactions and beam-type collisional activation of the polypeptide ions are performed in rapid succession in the high-pressure collision cell (Q2) of a quadrupole/time-of-flight tandem mass spectrometer (QqTOF), where the electron-transfer reagent anions are accumulated. Duty cycles for both electron-transfer dissociation (ETD) and collision-induced dissociation (CID) experiments are improved relative to ion trapping approaches since there are no discrete ion storage and reaction steps for ETD experiments and no discrete ion storage step and frequency tuning for CID experiments. For this technique, moderately high resolution and mass accuracy are also obtained due to mass analysis via the TOF analyzer. This relatively simple approach has been demonstrated with a triply charged tryptic peptide, a triply charged tryptic phosphopeptide, and a triply charged tryptic N-linked glycopeptide. For the tryptic peptide, the sequence is identified with more certainty than would be available from a single method alone due to the complementary information provided by these two dissociation methods. Because of the complementary information derived from both ETD and CID dissociation methods, peptide sequence and post-translational modification (PTM) sites for the phosphopeptide are identified. This combined ETD and CID approach is particularly useful for characterizing glycopeptides because ETD generates information about both peptide sequence and locations of the glycosylation sites, whereas CID provides information about the glycan structure.
Han
Hongling
H
Department of Chemistry, Purdue University, West Lafayette, Indiana 47907-2084, USA.
Xia
Yu
Y
Yang
Min
M
McLuckey
Scott A
SA
D000818:Animals; D002364:Caseins / Q000737:chemistry; D002417:Cattle; D006020:Glycopeptides / Q000737:chemistry; D006736:Horses; D009211:Myoglobin / Q000737:chemistry; D010455:Peptides / Q000737:chemistry*; D037121:Plant Lectins / Q000737:chemistry; D053719:Tandem Mass Spectrometry / Q000379:methods*; D014108:Tosylphenylalanyl Chloromethyl Ketone / Q000737:chemistry; D014357:Trypsin / Q000737:chemistry
D002364:Caseins; D006020:Glycopeptides; D009211:Myoglobin; D010455:Peptides; D037121:Plant Lectins; C045629:erythrina lectin; D014108:Tosylphenylalanyl Chloromethyl Ketone; D014357:Trypsin