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Proteomics Analysis of Contact-Initiated Eph Receptor–Ephrin Signaling

  • Claus Jorgensen
  • Alexei Poliakov
Protocol
  • 2.7k Downloads
Part of the Methods in Molecular Biology book series (MIMB, volume 1066)

Abstract

Large-scale biochemical analysis of cell-specific signaling can be interrogated in cocultures of Eph receptor- and ephrin-expressing cells by combining proteomics analysis with cell-specific metabolic labeling. In this chapter, we describe how to perform such large-scale analysis, including the generation of cells stably expressing the receptors and ligands of interest, optimization steps for Eph–ephrin coculture, and the proteomics analysis. As the experimental details may vary depending on the specific system that is being interrogated, the goal of the chapter is mainly to provide sufficient experimental context for experienced researchers to set up and conduct these experiments.

Key words

Proteomics Eph receptors Ephrins Cell–cell interactions Cell signaling 

Notes

Acknowledgements

The authors would like to acknowledge the contribution of Tony Pawson and David Wilkinson. Claus Jorgensen is a recipient of a CR-UK Career Establishment Award.

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Copyright information

© Springer Science+Business Media, New York 2013

Authors and Affiliations

  • Claus Jorgensen
    • 1
  • Alexei Poliakov
    • 2
  1. 1.Division of Cancer BiologyThe Institute of Cancer ResearchLondonUK
  2. 2.Division of Developmental NeurobiologyMRC National Institute of Medical ResearchLondonUK

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