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Complete protein characterization using top-down mass spectrometry and ultraviolet photodissociation.

ABSTRACT: 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.

SUBMITTER: Shaw JB

PROVIDER: S-EPMC3757099 | biostudies-literature | 2013 Aug

REPOSITORIES: biostudies-literature

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Complete protein characterization using top-down mass spectrometry and ultraviolet photodissociation.

Shaw Jared B JB Li Wenzong W Holden Dustin D DD Zhang Yan Y Griep-Raming Jens J Fellers Ryan T RT Early Bryan P BP Thomas Paul M PM Kelleher Neil L NL Brodbelt Jennifer S JS

Journal of the American Chemical Society 20130604 34

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 ...[more]

PMID: 23697802

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