Chapter 5. Production of Recombinant G Protein-Coupled Receptor in Yeast for Structural and Functional Analysis

  1. David R. Poyner1 and
  2. Mark Wheatley2
  1. Richard A. J. Darby,
  2. Mohammed Jamshad,
  3. Ljuban Grgic,
  4. William J. Holmes and
  5. Roslyn M. Bill

Published Online: 25 JAN 2010

DOI: 10.1002/9780470749210.ch5

G Protein-Coupled Receptors: Essential Methods

G Protein-Coupled Receptors: Essential Methods

How to Cite

Darby, R. A. J., Jamshad, M., Grgic, L., Holmes, W. J. and Bill, R. M. (2010) Production of Recombinant G Protein-Coupled Receptor in Yeast for Structural and Functional Analysis, in G Protein-Coupled Receptors: Essential Methods (eds D. R. Poyner and M. Wheatley), Wiley-Blackwell, Oxford, UK. doi: 10.1002/9780470749210.ch5

Editor Information

  1. 1

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

  2. 2

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

Author Information

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

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:

  • recombinant G protein-coupled receptor production in yeast - structural and functional analysis;
  • G protein-coupled receptors (GPCRs) - naturally present in native membranes;
  • preparation of expression plasmids;
  • DNA sequence encoding GPCR of interest - amplified via polymerase chain reaction (PCR);
  • PCR amplification and cloning;
  • Protocol 5.2 yeast transformations;
  • Protocol 5.3 screening for GPCR production in P. pastoris clones;
  • solubilization and purification;
  • Protocol 5.5 GPCR solubilization and purification;
  • analytical ultracentrifugation (AUC), technique characterizing solution state of macromolecules

Summary

This chapter contains sections titled:

  • Introduction

  • Methods and approaches

  • Troubleshooting

  • References