Implementation of Electron-Transfer Dissociation on a Hybrid Linear Ion Trap−Orbitrap Mass Spectrometer
- 19 April 2007
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 79 (10), 3525-3534
- https://doi.org/10.1021/ac070020k
Abstract
We describe the adaptation of a hybrid quadrupole linear ion trap−orbitrap mass spectrometer to accommodate electron-transfer ion/ion reactions (ETD) for peptide and protein characterization. The method utilizes pulsed, dual electrospray ion sources and requires minimal instrument modification. Switching between cation and reagent anion injection schemes is automated and accomplished within a few hundred milliseconds. Ion/ion reactions are conducted within the linear ion trap, after which the c- and z-type product ions are passed to the orbitrap for high-resolution m/z analysis. With this arrangement, mass accuracies are typically measured to within 2 ppm at a resolving power of ∼60 000. Using large peptides and intact proteins, we demonstrate such capabilities will accelerate our ability to interrogate high-mass species. To illustrate compatibility with automated data-dependent analysis and subsequent data processing, we couple the technique with an online chromatographic separation of a yeast whole-cell lysate followed by peptide identification using ProSight PC. Fairly long pulsing times and relatively low ET efficiency, as compared to conventional ETD instrumentation, are the main drawbacks of this approach. Still, our results suggest that the implementation of ETD on sensitive, high-resolution, and high-mass accuracy hybrid instrumentation, such as the orbitrap, will substantially propel the emergent fields of middle- and top-down proteomics.Keywords
This publication has 29 references indexed in Scilit:
- Supplemental Activation Method for High-Efficiency Electron-Transfer Dissociation of Doubly Protonated Peptide PrecursorsAnalytical Chemistry, 2006
- Electron-Transfer Reagent Anion Formation via Electrospray Ionization and Collision-Induced DissociationAnalytical Chemistry, 2006
- Implementation of Ion/Ion Reactions in a Quadrupole/Time-of-Flight Tandem Mass SpectrometerAnalytical Chemistry, 2006
- Phosphopeptide Anion Characterization via Sequential Charge Inversion and Electron-Transfer DissociationAnalytical Chemistry, 2006
- Alternately Pulsed Nanoelectrospray Ionization/Atmospheric Pressure Chemical Ionization for Ion/Ion Reactions in an Electrodynamic Ion TrapAnalytical Chemistry, 2006
- Proteomics-Grade de Novo Sequencing ApproachJournal of Proteome Research, 2005
- Ion/ion chemistry of high-mass multiply charged ionsMass Spectrometry Reviews, 1998
- Adaptation of the Paul trap for study of the reaction of multiply charged cations with singly charged anionsInternational Journal of Mass Spectrometry and Ion Processes, 1997
- Product Ion Charge State Determination via Ion/Ion Proton Transfer ReactionsAnalytical Chemistry, 1996
- Reflecting time-of-flight mass spectrometer with an electrospray ion source and orthogonal extractionAnalytical Chemistry, 1994