The use of site-directed mutagenesis to study GPCRs

Alex C. Conner, James Barwell, David R. Poyner, Mark Wheatley

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

G protein coupled receptors (GPCRs) are highly flexible and dynamic proteins, which are able to interact with diverse ligands, effectors, and regulatory proteins. Site-directed mutagenesis (SDM) is a powerful tool for providing insight into how these proteins actually work, both in its own right and when used in conjunction with information provided by other techniques such as crystallography or molecular modelling. Mutagenesis has been used to identify and characterise a myriad of functionally important residues, motifs and domains within the GPCR architecture, and to identify aspects of similarity and differences between the major families of GPCRs. This chapter presents the necessary information for undertaking informative SDM of these proteins. Whilst this is relevant to protein structure/function studies in general, specific pitfalls and protocols suited to investigating GPCRs in particular will be highlighted.

Original languageEnglish
Pages (from-to)85-98
Number of pages14
JournalMethods in Molecular Biology
Volume746
DOIs
Publication statusE-pub ahead of print - 5 May 2011
Externally publishedYes

Keywords

  • Alanine scan
  • Chimeric mutagenesis
  • Plasmid amplification
  • Plasmid transfection
  • Primer design
  • Random mutagenesis
  • Substituted cysteine accessibility method

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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