Top-down proteomics with a quadrupole time-of-flight mass spectrometer and collision-induced dissociation

Rapid Commun Mass Spectrom. 2009 Mar;23(5):661-6. doi: 10.1002/rcm.3925.

Abstract

With slight modifications of the instrumental parameters, we demonstrate that satisfactory top-down data can be obtained with collision-induced dissociation (CID) tandem mass spectrometry on a quadrupole time-of-flight (qTOF) instrument not originally designed for this purpose. Protein identification is achieved with both N- and C-terminal sequence tags and BLAST database searches. The accurate mass measurement of multiply charged fragment ions (mostly y and b-type) supplements the limited set of cleavage sites and provides a high degree of sequence coverage (90-100%). Post-translational modification issues can be addressed too. This approach might help those mass spectrometry (MS) core facilities that are not able to afford very high-resolution instruments, thus expanding the benefits of top-down protein analysis over the worldwide MS community.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Carbonic Anhydrase II / analysis
  • Carbonic Anhydrase II / chemistry*
  • Cattle
  • Gene Expression Profiling / methods*
  • Horses
  • Molecular Sequence Data
  • Myoglobin / analysis
  • Myoglobin / chemistry*
  • Proteome / analysis
  • Proteome / chemistry*
  • Proteomics / methods*
  • Spectrometry, Mass, Electrospray Ionization / methods*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*

Substances

  • Myoglobin
  • Proteome
  • Carbonic Anhydrase II