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 Citations (Scopus)

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 publicationAnnals of the New York Academy of Sciences
Pages21-27
Number of pages7
Volume1144
DOIs
StatePublished - Nov 2008

Publication series

NameAnnals of the New York Academy of Sciences
Volume1144
ISSN (Print)00778923
ISSN (Electronic)17496632

Fingerprint

Cholecystokinin Receptors
Oligomerization
Fluorescence Polarization
Occupations
Anisotropy
Fluorescence
Cells
Bioluminescence
Ligands
Fluorescence Resonance Energy Transfer
Monitoring
G-Protein-Coupled Receptors
Proteins

Keywords

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

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

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 Annals of the New York Academy of Sciences (Vol. 1144, pp. 21-27). (Annals of the New York Academy of Sciences; Vol. 1144). https://doi.org/10.1196/annals.1418.004

Monitoring the state of cholecystokinin receptor oligomerization after ligand binding using decay of time-resolved fluorescence anisotropy. / Harikumar, Kaleeckal G.; Miller, Laurence J.

Annals of the New York Academy of Sciences. Vol. 1144 2008. p. 21-27 (Annals of the New York Academy of Sciences; Vol. 1144).

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

Harikumar, KG & Miller, LJ 2008, Monitoring the state of cholecystokinin receptor oligomerization after ligand binding using decay of time-resolved fluorescence anisotropy. in Annals of the New York Academy of Sciences. vol. 1144, Annals of the New York Academy of Sciences, vol. 1144, pp. 21-27. https://doi.org/10.1196/annals.1418.004
Harikumar KG, Miller LJ. Monitoring the state of cholecystokinin receptor oligomerization after ligand binding using decay of time-resolved fluorescence anisotropy. In Annals of the New York Academy of Sciences. Vol. 1144. 2008. p. 21-27. (Annals of the New York Academy of Sciences). https://doi.org/10.1196/annals.1418.004
Harikumar, Kaleeckal G. ; Miller, Laurence J. / Monitoring the state of cholecystokinin receptor oligomerization after ligand binding using decay of time-resolved fluorescence anisotropy. Annals of the New York Academy of Sciences. Vol. 1144 2008. pp. 21-27 (Annals of the New York Academy of Sciences).
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