Chapter 8. A Disulfide Cross-linking Strategy Useful for Studying Ligand-induced Structural Changes in GPCRs

  1. David R. Poyner2 and
  2. Mark Wheatley3
  1. Jian Hua Li,
  2. Stuart D. C. Ward,
  3. Sung-Jun Han,
  4. Fadi F. Hamdan and
  5. Jürgen Wess

Published Online: 25 JAN 2010

DOI: 10.1002/9780470749210.ch8

G Protein-Coupled Receptors: Essential Methods

G Protein-Coupled Receptors: Essential Methods

How to Cite

Li, J. H., Ward, S. D. C., Han, S.-J., Hamdan, F. F. and Wess, J. (2010) A Disulfide Cross-linking Strategy Useful for Studying Ligand-induced Structural Changes in GPCRs, in G Protein-Coupled Receptors: Essential Methods (eds D. R. Poyner and M. Wheatley), Wiley-Blackwell, Oxford, UK. doi: 10.1002/9780470749210.ch8

Editor Information

  1. 2

    Pharmaceutical and Biological Sciences, School of Life and Health Sciences, Aston University, Birmingham B4 7ET, UK

  2. 3

    School of Biosciences, Birmingham University, Birmingham B15 2TT, UK

Author Information

  1. Molecular Signaling Section, Laboratory of Bioorganic Chemistry, National Institute of Diabetes and Digestive and Kidney Diseases, Bethesda, MD 20892, USA

Publication History

  1. Published Online: 25 JAN 2010
  2. Published Print: 22 JAN 2010

ISBN Information

Print ISBN: 9780470749142

Online ISBN: 9780470749210

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Keywords:

  • disulfide cross-linking strategy - studying ligand-induced structural changes in GPCRs;
  • G protein-coupled receptors (GPCRs) - largest group of cell-surface receptors found in nature;
  • monitoring ligand-induced changes in M3 mAChR structure;
  • rendering M3 mAChR in disulfide cross-linking studies;
  • agonist-promoted disulfide cross-link formation in double Cys mutant M3 mAChRs;
  • agonist-induced conformational changes;
  • cell culture, transfection and membrane preparation;
  • specific double Cys mutant receptor forming intramolecular disulfide bond;
  • Protocol 8.3 urea treatment of receptor-containing membranes;
  • Protocol 8.6 Western blotting analysis

Summary

This chapter contains sections titled:

  • Introduction

  • Methods and approaches

  • General considerations, caveats and troubleshooting

  • Acknowledgements

  • References