Time-Resolved Förster Resonance Energy Transfer-Based Technologies to Investigate G Protein-Coupled Receptor Machinery: High-Throughput Screening Assays and Future Development - Progress in Molecular Biology and Translational Science - page 285

Table 7.1 Summary of TR-FRET-compatible assays for GPCR
GPCR signaling
cascade and
downstream events

Biological
measurement

Available TR-FRET assay
(company)

Illustrative
paper

Basis

Zwier et al. 19 Ligand binding Displacement of a

reference ligand

Tag-lite (Cisbio Bioassays) Ligand binding and/or displacement of a reference

fluorescent ligand that generates TR-FRET
signal with a Lumi4-Tb labeled SNAP-tagged
GPCR

Handl et al. 20 DELFIA receptor-ligand-

binding assays (Perkin Elmer)

An europium-labeled ligand is competed off with
a tested unlabeled ligand. The amount of
specifically bound labeled ligand is revealed by
TR-fluorescence after addition of an
enhancement solution and wash steps to remove
unbound ligand

Frang et al. 15 G protein-dependent

events

Guanine nucleotide
exchange

DELFIA Eu-GTP binding
assay (Perkin Elmer)

Binding of a nonhydrolyzable europium-
labeled GTP analog to receptor-activated G
proteins

Gq pathway Inositol

monophosphate
(IP1) accumulation

IP-One (Cisbio Bioassays) Competition assay between endogenously

produced IP1 and acceptor-labeled IP1 for
binding on donor-labeled IP1 antibody
(TR-FRET signal decreases as endogenous
IP1 is produced)

Trinquet
et al. 21
Trinquet
et al. 22