TY - JOUR
T1 - Using Bioluminescence Resonance Energy Transfer (BRET) to Characterize Agonist-Induced Arrestin Recruitment to Modified and Unmodified G Protein-Coupled Receptors
AU - Donthamsetti, Prashant
AU - Quejada, Jose Rafael
AU - Javitch, Jonathan A.
AU - Gurevich, Vsevolod V.
AU - Lambert, Nevin A.
N1 - Funding Information:
Supported by NIH grants GM077561, GM109955, EY011500 (VVG), NIMH-R01 MH54137 (PCD, JRQ, JAJ), DA022413 (JAJ), GM078319 (NAL).
Publisher Copyright:
Copyright © 2015 John Wiley & Sons, Inc.
PY - 2015/9/1
Y1 - 2015/9/1
N2 - G protein-coupled receptors (GPCRs) represent ∼25% of current drug targets. Ligand binding to these receptors activates G proteins and arrestins, which are involved in differential signaling pathways. Because functionally selective or biased ligands activate one of these two pathways, they may be superior medications for certain diseases states. The identification of such ligands requires robust drug screening assays for both G protein and arrestin activity. This unit describes protocols for two bioluminescence resonance energy transfer (BRET)-based assays used to monitor arrestin recruitment to GPCRs. One assay requires modification of GPCRs by fusion to a BRET donor or acceptor moiety, whereas the other can detect arrestin recruitment to unmodified GPCRs.
AB - G protein-coupled receptors (GPCRs) represent ∼25% of current drug targets. Ligand binding to these receptors activates G proteins and arrestins, which are involved in differential signaling pathways. Because functionally selective or biased ligands activate one of these two pathways, they may be superior medications for certain diseases states. The identification of such ligands requires robust drug screening assays for both G protein and arrestin activity. This unit describes protocols for two bioluminescence resonance energy transfer (BRET)-based assays used to monitor arrestin recruitment to GPCRs. One assay requires modification of GPCRs by fusion to a BRET donor or acceptor moiety, whereas the other can detect arrestin recruitment to unmodified GPCRs.
KW - G protein-coupled receptors (GPCRs)
KW - arrestin
KW - bioluminescence resonance energy transfer (BRET)
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U2 - 10.1002/0471141755.ph0214s70
DO - 10.1002/0471141755.ph0214s70
M3 - Article
C2 - 26331887
AN - SCOPUS:85010648012
SN - 1934-8282
VL - 70
SP - 2.14.1-2.14.14
JO - Current protocols in pharmacology
JF - Current protocols in pharmacology
IS - 1
ER -