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Related Experiment Videos

Proteomic complex detection using sedimentation.

Nicholas T Hartman1, Francesca Sicilia, Kathryn S Lilley

  • 1Department of Biochemistry, University of Cambridge, Building O, Downing Site, Cambridge CB2 1QW, United Kingdom.

Analytical Chemistry
|March 1, 2007
PubMed
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We developed Proteomic Complex Detection using Sedimentation (ProCoDeS), a new method to identify protein complexes by analyzing how proteins sediment. This technique helps discover new protein interactions within cells.

Area of Science:

  • Proteomics
  • Biochemistry
  • Molecular Biology

Background:

  • Protein-protein interactions are crucial for cellular functions.
  • Existing methods for screening novel protein complexes are limited.
  • Understanding protein complex composition is vital for deciphering cellular mechanisms.

Purpose of the Study:

  • To introduce a novel quantitative proteomics technique, ProCoDeS (Proteomic Complex Detection using Sedimentation).
  • To profile the sedimentation behavior of numerous proteins within a rate zonal centrifugation gradient.
  • To enable the identification of proteins within stable protein complexes.

Main Methods:

  • Proteomic Complex Detection using Sedimentation (ProCoDeS) involves rate zonal centrifugation.
  • Quantitative analysis of protein abundance across gradient fractions using ICAT labeling and electrospray mass spectrometry.

Related Experiment Videos

  • Profiling sedimentation of solubilized mitochondrial membrane proteins from Arabidopsis thaliana.

    • Main Results:

    • Proteins within the same complex exhibited highly similar sedimentation profiles, confirming method reproducibility.
    • Proteins identified in complexes sedimented faster than their predicted monomer molecular weights.
    • The sedimentation rate relative to known complexes allowed for approximate size inference of unknown complexes.

    Conclusions:

    • ProCoDeS is an effective quantitative proteomics technique for identifying protein complexes.
    • The method allows for the screening of various biological samples (tissues, cells, organelles).
    • ProCoDeS facilitates the discovery of stable protein complexes for further characterization.