TY - GEN
T1 - Monitoring the state of cholecystokinin receptor oligomerization after ligand binding using decay of time-resolved fluorescence anisotropy
AU - Harikumar, Kaleeckal G.
AU - Miller, Laurence J.
PY - 2008/11
Y1 - 2008/11
N2 - 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.
AB - 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.
KW - Cholecystokinin receptor
KW - G protein-coupled receptors
KW - Receptor oligomerization
KW - Rotational dynamics
KW - Time-resolved anisotropy
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U2 - 10.1196/annals.1418.004
DO - 10.1196/annals.1418.004
M3 - Conference contribution
C2 - 19076359
AN - SCOPUS:56649109884
SN - 9781573317047
T3 - Annals of the New York Academy of Sciences
SP - 21
EP - 27
BT - Neural Signaling Opportunities for Novel Diagnostic Approaches and Therapies
PB - Blackwell Publishing Inc.
ER -