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Related Experiment Videos

Simultaneous quantification and identification using 18O labeling with an ion trap mass spectrometer and the analysis

Wayne A Hicks1, Brian D Halligan, Ronit Y Slyper

  • 1National Proteomics Research Center, Medical College of Wisconsin, 8701 Watertown Plank Road, Milwaukee, WI 53213, USA. whicks@mcw.edu

Journal of the American Society for Mass Spectrometry
|May 24, 2005
PubMed
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Stable isotope labeling with oxygen-18 ((18)O) enables quantitative proteomics. New methods using zoom scans and ZoomQuant software improve (18)O labeling accuracy with electrospray ionization-ion trap mass spectrometry.

Area of Science:

  • Proteomics
  • Analytical Chemistry
  • Biochemistry

Background:

  • Stable isotope labeling with oxygen-18 ((18)O) offers combined qualitative and quantitative protein expression analysis.
  • Limited automated quantification methods and small mass shifts hinder its application with electrospray ionization-ion trap (ESI-IT) mass spectrometers.

Purpose of the Study:

  • To develop and validate a method for accurate (18)O labeling quantification using ESI-IT mass spectrometry.
  • To enable automated analysis of large datasets for differential protein expression studies.

Main Methods:

  • Utilized zoom scans for high-resolution ESI-IT mass spectrometric data acquisition.
  • Employed the ZoomQuant software tool for automated analysis of large datasets.
  • Calculated (18)O/(16)O ratios for differentially labeled peptides.

Related Experiment Videos

Main Results:

  • Achieved accurate calculation of relative peptide ratios even with 50% singly labeled species.
  • Obtained sufficient resolution for accurate ratio calculation in peptides with +3 charge states and lower.
  • Demonstrated isotopic ratio accuracy of approximately 21% using zoom scan data and ZoomQuant.
  • Reported sequence coverage comparable to standard MS and MS/MS acquisition modes.

Conclusions:

  • The combined approach of zoom scan data acquisition and ZoomQuant analysis significantly enhances the utility of (18)O labeling with ESI-IT mass spectrometry.
  • This method provides accurate and automated quantification for differential protein expression studies, comparable to other high-resolution mass spectrometry techniques.