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Related Experiment Video

Updated: Jun 3, 2026

Utilizing a Comprehensive Immunoprecipitation Enrichment System to Identify an Endogenous Post-translational Modification Profile for Target Proteins
08:12

Utilizing a Comprehensive Immunoprecipitation Enrichment System to Identify an Endogenous Post-translational Modification Profile for Target Proteins

Published on: January 8, 2018

A differential proteome screening system for post-translational modification-dependent transcription factor

Ole Pless1, Elisabeth Kowenz-Leutz, Gunnar Dittmar

  • 1Max-Delbrück-Center for Molecular Medicine, Berlin, Germany.

Nature Protocols
|March 5, 2011
PubMed
Summary

Array peptide screening (APS) identifies proteins interacting with modified transcription factors. This method reveals post-translational modification-dependent interactions, offering a cost-effective alternative to mass spectrometry.

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Area of Science:

  • Molecular Biology
  • Proteomics
  • Biochemistry

Background:

  • Post-translational modifications (PTMs) regulate transcription factor (TF) interactions with co-regulators and epigenetic modifiers.
  • Arginine and lysine methylation in conserved domains of C/EBP transcription factors suggests modification-dependent cofactor docking.

Purpose of the Study:

  • To introduce and validate Array Peptide Screening (APS) for detecting PTM-dependent protein interactions.
  • To demonstrate APS as a systematic, differential, and cost-effective method for proteome-wide interaction discovery.

Main Methods:

  • Chemically synthesized, biotinylated peptides (modified and non-modified) coupled to fluorophore-labeled streptavidin.
  • Parallel application of peptides to bacterial expression libraries in an arrayed format.

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Last Updated: Jun 3, 2026

Utilizing a Comprehensive Immunoprecipitation Enrichment System to Identify an Endogenous Post-translational Modification Profile for Target Proteins
08:12

Utilizing a Comprehensive Immunoprecipitation Enrichment System to Identify an Endogenous Post-translational Modification Profile for Target Proteins

Published on: January 8, 2018

Identification of MyoD Interactome Using Tandem Affinity Purification Coupled to Mass Spectrometry
14:47

Identification of MyoD Interactome Using Tandem Affinity Purification Coupled to Mass Spectrometry

Published on: May 17, 2016

  • Detection and quantification of differential protein binding via laser scanning.

    • Main Results:

    • APS successfully identifies proteins that differentially bind to modified versus non-modified peptides.
    • The method was previously applied to study arginine methylation effects on C/EBPβ peptides.
    • APS allows determination of PTM-dependent TF interactions, relative binding affinities, and rapid protein classification.

    Conclusions:

    • APS is a robust method for discovering PTM-dependent protein interactions, independent of peptide transactivation potential or protein abundance.
    • This technique offers a rapid (3-4 days) and cost-effective alternative to mass spectrometry for interaction studies.