Unit

UNIT 19.21 Identification of Protein-Protein Interactions by Surface Plasmon Resonance followed by Mass Spectrometry

  1. Alexandra Madeira1,
  2. Elisabet Vikeved1,
  3. Anna Nilsson2,
  4. Benita Sjögren1,
  5. Per E. Andrén2,
  6. Per Svenningsson1

Published Online: 1 AUG 2011

DOI: 10.1002/0471140864.ps1921s65

Current Protocols in Protein Science

Current Protocols in Protein Science

How to Cite

Madeira, A., Vikeved, E., Nilsson, A., Sjögren, B., Andrén, P. E. and Svenningsson, P. 2011. Identification of Protein-Protein Interactions by Surface Plasmon Resonance followed by Mass Spectrometry. Current Protocols in Protein Science. 65:19.21:19.21.1–19.21.9.

Author Information

  1. 1

    Karolinska Institute, Stockholm, Sweden

  2. 2

    Uppsala University, Biomedical Centre, Uppsala, Sweden

Publication History

  1. Published Online: 1 AUG 2011
  2. Published Print: AUG 2011

Abstract

Elucidation of the function and meaning of the protein networks can be useful in the understanding of many pathological processes and the identification of new therapeutic targets. This unit describes an approach to discover protein-protein interactions by coupling surface plasmon resonance to mass spectrometry. Briefly, a protein is covalently bound to a sensor chip, which is then exposed to brain extracts injected over the surface via a microfluidic system. This allows the monitoring in real-time of the interactions between the immobilized ligand and the extracts. Interacting proteins from the extracts are then recovered, trypsinized, and identified using mass spectrometry. The data obtained are searched against a sequence database using the Mascot software. To exclude nonspecific interactors, control experiments using blank sensor chips, and/or randomized peptides, are performed. The protocol presented here does not require specific labeling or modification of proteins and can be performed in <4 days. Curr. Protoc. Protein Sci. 65:19.21.1-19.21.9. © 2011 by John Wiley & Sons, Inc.

Keywords:

  • SPR;
  • mass spectrometry;
  • protein interactions