Journal
JOURNAL OF PROTEOME RESEARCH
Volume 7, Issue 8, Pages 3127-3136Publisher
AMER CHEMICAL SOC
DOI: 10.1021/pr800264t
Keywords
electron transfer dissociation; ion-ion; proteomics; tandem mass spectrometry; top-down; orbitrap
Categories
Funding
- Division Of Chemistry
- Direct For Mathematical & Physical Scien [0747990] Funding Source: National Science Foundation
- NHGRI NIH HHS [T32 HG002760-02, T32 HG002760-05, T32 HG002760-01, T32 HG002760-04, T32 HG002760-03, T32 HG002760, 5 T32 HG 002706] Funding Source: Medline
- NIGMS NIH HHS [R01 GM080148, T32 GM008349-19, 1 R01 GM 080148, T32 GM008349-10, T32 GM008349-12, T32 GM008349-11, T32 GM008349-15S1, T32 GM008349-14, T32 GM008349, 5 T32 GM 08349, T32 GM008349-15, T32 GM008349-17, T32 GM008349-18, R01 GM080148-01A1, T32 GM008349-16, T32 GM008349-13] Funding Source: Medline
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Here we detail the modification of a quadrupole linear ion trap-orbitrap hybrid (QLT-orbitrap) mass spectrometer to accommodate a negative chemical ionization (NCI) source. The NO source is used to produce fluoranthene radical anions for imparting electron transfer dissociation (ETD). The anion beam is stable, robust, and intense so that a sufficient amount of reagents can be injected into the QLT in only 4-8 ms. Following ion/ion reaction in the QLT, ETD product ions are mass-to-charge (m/z) analyzed in either the QLT (for speed and sensitivity) or the orbitrap (for mass resolution and accuracy). Here we describe the physical layout of this device, parametric optimization of anion transport, an evaluation of relevant ETD figures of merit, and the application of this instrument to protein sequence analysis. Described proteomic applications include complex peptide mixture analysis, post-translational modification (PTM) site identification, isotope-encoded quantitation, large peptide characterization, and intact protein analysis. From these experiments, we conclude the ETD-enabled orbitrap will provide the proteomic field with several new opportunities and represents an advance in protein sequence analysis technologies.
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