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Two-Dimensional Mass Spectrometry for Proteomics, a Comparative Study with Cytochrome c.
Maria A van Agthoven1, Christopher A Wootton1, Lionel Chiron2
1Department of Chemistry, University of Warwick , Gibbet Hill Road, CV4 7AL Coventry, West Midlands, United Kingdom.
Analytical Chemistry
|March 19, 2016
Summary
Two-dimensional Fourier transform ion cyclotron resonance mass spectrometry (2D FT-ICR MS) offers accurate precursor-fragment ion correlation without prior isolation. This data-independent method enhances structural analysis in complex samples with improved accuracy over traditional MS/MS.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Mass Spectrometry
Background:
- Tandem mass spectrometry (MS/MS) typically requires precursor ion isolation, limiting analysis of complex mixtures.
- Two-dimensional Fourier transform ion cyclotron resonance mass spectrometry (2D FT-ICR MS) enables precursor-fragment ion correlation without prior isolation.
- Optimization of 2D FT-ICR MS as a data-independent method is crucial for structural analysis.
Purpose of the Study:
- To explore the capabilities of 2D FT-ICR MS for structural analysis of complex samples.
- To evaluate 2D FT-ICR MS using ECD and IRMPD fragmentation on a tryptic digest of cytochrome c.
- To compare the accuracy and efficiency of 2D FT-ICR MS with traditional MS/MS methods.
Main Methods:
- Utilized two-dimensional Fourier transform ion cyclotron resonance mass spectrometry (2D FT-ICR MS).
- Employed both electron capture dissociation (ECD) and infrared multiphoton dissociation (IRMPD) fragmentation techniques.
- Applied quadratic calibration for fragment ion peak assignment and analyzed a tryptic digest of cytochrome c.
Main Results:
- 2D FT-ICR MS provided useful peptide fragmentation patterns over a dynamic range of nearly 400.
- Fragment ion assignment was successful, even for precursors differing by only m/z 0.0156.
- Correlation accuracy between precursor and fragment ions in 2D FT-ICR MS surpassed quadrupole-isolated MS/MS spectra.
- Achieved cleavage coverage for cytochrome c comparable to high-resolution FT-ICR MS/MS but with a single experimental scan.
Conclusions:
- 2D FT-ICR MS is a powerful, data-independent analytical tool for structural elucidation in complex samples.
- The method offers superior accuracy and efficiency compared to traditional MS/MS approaches, particularly for closely related ions.
- Single-scan 2D FT-ICR MS analysis provides comprehensive fragmentation data, approaching the quality of multi-scan high-resolution MS/MS.
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