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

J Am Chem Soc. 2013 Aug 28;135(34):12646-51. doi: 10.1021/ja4029654. Epub 2013 Jun 4.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Humans
  • Mass Spectrometry
  • Models, Molecular
  • NIMA-Interacting Peptidylprolyl Isomerase
  • Peptidylprolyl Isomerase / analysis*
  • Peptidylprolyl Isomerase / genetics
  • Photochemical Processes
  • Proteomics*
  • Ultraviolet Rays*

Substances

  • NIMA-Interacting Peptidylprolyl Isomerase
  • PIN1 protein, human
  • Peptidylprolyl Isomerase