Functional GPCR microarrays.
This paper describes G-protein-coupled receptor (GPCR) microarrays on porous glass substrates and functional assays based on the binding of a europium-labeled GTP analogue. The porous glass slides were made by casting a glass frit on impermeable glass slides and then coating with gamma-aminopropyl silane (GAPS). The emitted fluorescence was captured on an imager with a time-gated intensified CCD detector. Microarrays of the neurotensin receptor 1, the cholinergic receptor muscarinic 2, the opioid receptor mu, and the cannabinoid receptor 1 were fabricated by pin printing. The selective agonism of each of the receptors was observed. The screening of potential antagonists was demonstrated using a cocktail of agonists. The amount of activation observed was sufficient to permit determinations of EC50 and IC50. Such microarrays could potentially streamline drug discovery by helping integrate primary screening with selectivity and safety screening without compromising the essential functional information obtainable from cellular assays.
Hong
Yulong
Y
Science and Technology Division, Corning Inc., Corning, NY 14831, USA.
Webb
Brian L
BL
Su
Hui
H
Mozdy
Eric J
EJ
Fang
Ye
Y
Wu
Qi
Q
Liu
Li
L
Beck
Jonathan
J
Ferrie
Ann M
AM
Raghavan
Srikanth
S
Mauro
John
J
Carre
Alain
A
Müeller
Dirk
D
Lai
Fang
F
Rasnow
Brian
B
Johnson
Michael
M
Min
Hosung
H
Salon
John
J
Lahiri
Joydeep
J
D004353:Drug Evaluation, Preclinical / Q000379:methods; D005456:Fluorescent Dyes; D008024:Ligands; D040081:Protein Array Analysis / Q000295:instrumentation / Q000379:methods*; D043562:Receptors, G-Protein-Coupled / Q000037:antagonists & inhibitors*
D005456:Fluorescent Dyes; D008024:Ligands; D043562:Receptors, G-Protein-Coupled