Journal of the American Chemical Society · 2013 · 366 citations · 33 references
Life SciencesBiochemistryMolecular BiologyMass Spectrometric AnalysisProteomicsSpectroscopyBottom-up ProteomicsMass SpectrometryMass Spectrometry-based ProteomicsPost-translational Modification AnalysisProtein IdentificationTop-down ProteomicsActivation EnergyFragmentationMutationOrbitrap Mass AnalyzersOrbitrap Mass SpectrometryPhotodissociation SpectroscopyProtein Mass SpectrometryProtein Modifications
The top-down approach to proteomics offers compelling advantages due to the potential to provide complete characterization of protein sequence and post-translational modifications. Here we describe the implementation of 193 nm ultraviolet photodissociation (UVPD) in an Orbitrap mass spectrometer for characterization of intact proteins. Near-complete fragmentation of proteins up to 29 kDa is achieved with UVPD including the unambiguous localization of a single residue mutation and several protein modifications on Pin1 (Q13526), a protein implicated in the development of Alzheimer's disease and in cancer pathogenesis. The 5 ns, high-energy activation afforded by UVPD exhibits far less precursor ion-charge state dependence than conventional collision- and electron-based dissociation methods.
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