Abstract
A large variety of techniques has been used to monitor activation
of G protein-coupled receptors (GPCRs) both in isolated membranes
and in intact cells. However, most of these techniques cannot resolve
receptor activation and signaling in space and in time. Here, we
describe techniques that allow the temporally and spatially resolved
monitoring of these processes by optical recording with energy transfer
techniques. Fluorescence and bioluminescence resonance energy transfer,
FRET and BRET, are based on energy transfer between two closely spaced
probes. The exquisite sensitivity of FRET and BRET to the distance
of the two probes makes these techniques ideal tools to study either
protein-protein interactions (when the two probes are localized on
two different proteins) or conformational changes within a given
protein (when the two probes are localized on a single protein).
Here, we review the latter approach as a tool to study receptor activation
and the levels of the second messengers cAMP and cGMP in intact cells.
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