title: Two-Dimensional Mass Spectrometry for Proteomics, a Comparative Study with Cytochrome c.
abstract: 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.
authors.affiliation: Department of Chemistry, University of Warwick , Gibbet Hill Road, CV4 7AL Coventry, West Midlands, United Kingdom.
authors.forename: Maria A
authors.initials: MA
authors.lastname: van Agthoven
authors.affiliation: Department of Chemistry, University of Warwick , Gibbet Hill Road, CV4 7AL Coventry, West Midlands, United Kingdom.
authors.forename: Christopher A
authors.initials: CA
authors.lastname: Wootton
authors.affiliation: CASC4DE, Le Lodge, 20, av. du Neuhof, 67100 Strasbourg, France.
authors.forename: Lionel
authors.initials: L
authors.lastname: Chiron
authors.affiliation: NMRTEC, Bld. Sébastien Brandt, Bioparc - Bat. B, 67400 Illkirch-Graffenstaden, France.
authors.forename: Marie-Aude
authors.initials: MA
authors.lastname: Coutouly
authors.affiliation: Department of Chemistry, University of Warwick , Gibbet Hill Road, CV4 7AL Coventry, West Midlands, United Kingdom.
authors.forename: Andrew
authors.initials: A
authors.lastname: Soulby
authors.affiliation: Department of Chemistry, University of Warwick , Gibbet Hill Road, CV4 7AL Coventry, West Midlands, United Kingdom.
authors.forename: Juan
authors.initials: J
authors.lastname: Wei
authors.affiliation: Department of Chemistry, University of Warwick , Gibbet Hill Road, CV4 7AL Coventry, West Midlands, United Kingdom.
authors.forename: Mark P
authors.initials: MP
authors.lastname: Barrow
authors.affiliation: CASC4DE, Le Lodge, 20, av. du Neuhof, 67100 Strasbourg, France.|Institut de Génétique et de Biologie Moléculaire et Cellulaire, INSERM, U596; CNRS, UMR7104; Université de Strasbourg , 1 rue Laurent Fries, 67404 Illkirch-Graffenstaden, France.
authors.forename: Marc-André
authors.initials: MA
authors.lastname: Delsuc
authors.affiliation: Université de Lille, CNRS, USR 3290, MSAP, Miniaturisation pour la Synthèse l'Analyse et la Protéomique, FR 3688, FRABIO, Biochimie Structurale & Fonctionnelle des Assemblages Biomoléculaires, and FR 2638, Institut Eugène-Michel Chevreul, F-59000 Lille, France.
authors.forename: Christian
authors.initials: C
authors.lastname: Rolando
authors.affiliation: Department of Chemistry, University of Warwick , Gibbet Hill Road, CV4 7AL Coventry, West Midlands, United Kingdom.
authors.forename: Peter B
authors.initials: PB
authors.lastname: O'Connor
mesh_terms: D000465:Algorithms; D000818:Animals; D002417:Cattle; D045304:Cytochromes c / Q000032:analysis*; D005583:Fourier Analysis; D013058:Mass Spectrometry / Q000379:methods*; D040901:Proteomics / Q000379:methods*
chemical_list: D045304:Cytochromes c