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Abstract for Advances In Bioluminescent Assays for
G-Protein-Coupled Receptors
Frank Fan, Aileen Paguio, Braeden Butler, Brock Binkowski, Pete
Stecha and Keith Wood
Promega Corporation, 2800 Woods Hollow Rd Madison, WI 53711
Bioluminescent assays are uniquely suited for high throughput
screening due to their inherent high sensitivity, wide dynamic range
and low susceptibility to compound interference. G-protein-coupled
receptors constitute a major target class for the development of new
drugs. The activity of these receptors can be monitored via readouts
at various stages of the signal transduction cascade. Traditional
bioluminescent GPCR assay involves using luciferase as a genetic
reporter linked to appropriate response elements. Recent
improvements in reporter technology have enhanced the dynamics,
sensitivity, and reliability of these assays. A new assay allows
quantitation of cAMP in cell lysates through its regulation of
protein kinase A activity, where the ATPase activity can be detected
using bioluminescence. Another novel assay format based on a
genetically encoded luciferase biosensor allows direct detection of
cAMP within living cells. Comparison studies of these assays using
HEK293 cells expressing dopamine receptor D1 are presented.
Together, these bioluminescent assays provide more tools for GPCR
drug discovery. |
