Two-Dimensional Mass Spectrometry for Proteomics, a Comparative Study with Cytochrome c

• Published in: Analytical chemistry (Washington) Vol. 88; no. 8; pp. 4409 - 4417
• Main Authors: van Agthoven, Maria A, Wootton, Christopher A, Chiron, Lionel, Coutouly, Marie-Aude, Soulby, Andrew, Wei, Juan, Barrow, Mark P, Delsuc, Marc-André, Rolando, Christian, O’Connor, Peter B
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 19-04-2016
• Subjects: Algorithms; Analytical chemistry; Animals; Biochemistry, Molecular Biology; Cattle; Cytochrome; Cytochromes; Cytochromes c - analysis; Fourier Analysis; Fragmentation; Ions; Life Sciences; Mass spectra; Mass spectrometry; Mass Spectrometry - methods; Mass spectroscopy; Peptides; Precursors; Proteomics; Proteomics - methods; Quadrupoles; Two dimensional
• ISSN: 0003-2700; 1520-6882
• DOI: 10.1021/acs.analchem.5b04878

Two-dimensional Fourier transform ion cyclotron resonance mass spectrometry (2D FT-ICR MS) allows the correlation between precursor and fragment ions in tandem mass spectrometry without the need to isolate the precursor ion beforehand. 2D FT-ICR MS has been optimized as a data-independent method for the structural analysis of compounds in complex samples. Data processing methods and denoising algorithms have been developed to use it as an analytical tool. In the present study, the capabilities of 2D FT-ICR MS are explored with a tryptic digest of cytochrome c with both ECD and IRMPD as fragmentation modes. The 2D mass spectra showed useful fragmentation patterns of peptides over a dynamic range of almost 400. By using a quadratic calibration, fragment ion peaks could be successfully assigned. The correlation between precursor and fragment ions in the 2D mass spectra was more accurate than in MS/MS spectra after quadrupole isolation, due to the limitations of quadrupole isolation. The use of the second dimension allowed for successful fragment assignment from precursors that were separated by only m/z 0.0156. The resulting cleavage coverage of cytochrome c almost matched data provided by high-resolution FT-ICR MS/MS analysis, but the 2D FT-ICR MS method required only one experimental scan.