The use of site-directed mutagenesis to study GPCRs

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

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)

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
Title of host publicationReceptor signal transduction protocols
EditorsGary B. Willars, R.A. John Challiss
PublisherHumana Press
Pages85-98
Number of pages14
Edition3rd ed
ISBN (Electronic)978-1-61779-126-0
ISBN (Print)978-1-61779-125-3
DOIs
Publication statusPublished - 5 May 2011

Publication series

NameMethods in molecular biology
PublisherSpringer
Volume746
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • cultured cells
  • genetic vectors
  • site-directed mutagenesis
  • mutant chimeric proteins
  • G-protein-coupled receptors
  • transfection

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  • Cite this

    Conner, A. C., Barwell, J., Poyner, D. R., & Wheatley, M. (2011). The use of site-directed mutagenesis to study GPCRs. In G. B. Willars, & R. A. J. Challiss (Eds.), Receptor signal transduction protocols (3rd ed ed., pp. 85-98). (Methods in molecular biology; Vol. 746). Humana Press. https://doi.org/10.1007/978-1-61779-126-0_5