Protein quantification by selective isolation and fragmentation of isotopic pairs using FT-ICR MS

J Am Soc Mass Spectrom. 2008 Jul;19(7):973-7. doi: 10.1016/j.jasms.2008.03.012. Epub 2008 Apr 8.

Abstract

Isolation of tryptic peptide ions, along with their differentially labeled analogs derived from an artificial QconCAT protein, is performed using multiple correlated harmonic excitation fields in an FT-ICR cell. Simultaneous fragmentation of the isolated unlabeled and labeled peptide pairs using IRMPD yields specific y-series fragment ions useful for quantification. The mass increment attributed to stable isotope labeling at the C-terminus is maintained in the C-terminal fragment ions, providing multiple measurements of labeled/unlabeled intensity ratios during highly selective detection. The utility of this approach has been demonstrated in the absolute quantification of components of an unfractionated chicken muscle protein mixture.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Carbon Isotopes / chemistry
  • Chickens
  • Cyclotrons
  • Muscle, Skeletal / chemistry
  • Nitrogen Isotopes / chemistry
  • Peptides / chemistry*
  • Spectrometry, Mass, Electrospray Ionization
  • Spectroscopy, Fourier Transform Infrared / methods*
  • Tandem Mass Spectrometry
  • Trypsin / chemistry

Substances

  • Carbon Isotopes
  • Nitrogen Isotopes
  • Peptides
  • Trypsin