Monitoring the state of cholecystokinin receptor oligomerization after ligand binding using decay of time-resolved fluorescence anisotropy

Kaleeckal G. Harikumar, Laurence J. Miller

Research output: Chapter in Book/Report/Conference proceedingConference contribution

2 Scopus citations

Abstract

Oligomeric complexes of G protein-coupled receptors (GPCRs) are now commonly recognized and can provide a mechanism for regulation of signaling systems. Receptor oligomerization has been most extensively studied using coimmunoprecipitation and bioluminescence or fluorescence resonance energy-transfer techniques. Here, we have utilized decay of time-resolved fluorescence anisotropy of yellow fluorescent protein-labeled cholecystokinin receptor constructs to examine the state of oligomerization of this receptor in living cells. The rotational correlation times established that the cholecystokinin receptor is constitutively present in an oligomeric state that is dissociated in response to agonist occupation. In contrast, antagonist occupation failed to modify this signal, leaving the oligomeric structure intact. This dynamic technique complements the other biochemical and steady-state fluorescence techniques to establish the presence of oligomeric receptor complexes in living cells.

Original languageEnglish (US)
Title of host publicationNeural Signaling Opportunities for Novel Diagnostic Approaches and Therapies
PublisherBlackwell Publishing Inc.
Pages21-27
Number of pages7
ISBN (Print)9781573317047
DOIs
StatePublished - Nov 2008

Publication series

NameAnnals of the New York Academy of Sciences
Volume1144
ISSN (Print)0077-8923
ISSN (Electronic)1749-6632

Keywords

  • Cholecystokinin receptor
  • G protein-coupled receptors
  • Receptor oligomerization
  • Rotational dynamics
  • Time-resolved anisotropy

ASJC Scopus subject areas

  • Neuroscience(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • History and Philosophy of Science

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    Harikumar, K. G., & Miller, L. J. (2008). Monitoring the state of cholecystokinin receptor oligomerization after ligand binding using decay of time-resolved fluorescence anisotropy. In Neural Signaling Opportunities for Novel Diagnostic Approaches and Therapies (pp. 21-27). (Annals of the New York Academy of Sciences; Vol. 1144). Blackwell Publishing Inc.. https://doi.org/10.1196/annals.1418.004