Cell-based assays for G-protein-coupled receptor-mediated...

Details Cell-based assays for G-protein-coupled receptor-mediated... Cell-based assays for G-protein-coupled receptor-mediated...

2011-03-01

2011-03-01

Ulm, John D (Department: 1649)

Chemistry: molecular biology and microbiology

Animal cell, per se ; composition thereof; process of...

C435S007210, C435S069100

Reexamination Certificate

active

07897386

ABSTRACT:
Disclosed are compositions and methods for their use, such as in identifying G-protein-coupled receptors, ligands and compounds that modulate the activities of G-protein-coupled receptors. The compositions and methods employ cyclic nucleotide-gated channels and fluorescence dyes in detecting changes of intracellular cAMP levels in response to the stimulation of G-protein-coupled receptors. Activation of the G-protein-coupled receptors can be detected in a variety of assays, including cell-based imaging assays with fluorescence microscopes and high throughput assays with multi-well plates and fluorescence plate readers.

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Scott et al. Three Residues Predicted by Molecular Modeling to Interact with the Purine Moiety Alter Ligand Binding and Channel Gating in Cyclic Nucleotide-Gated Channels, 1998, Biochemistry 37:17239-17252.
Reinscheid, et al. High-throughput real-time monitoring of Gs-coupled receptor activation in intact cells using cyclic nucleotide-gated channels. Eur J Pharmacol. Sep. 30, 2003;478(1):27-34.
Nevin, et al. A pore-lining glutamic acid in the rat olfactory cyclic nucleotide-gated channel controls external spermine block. Neurosci Lett. Dec. 22, 2000;296(2-3):163-7.
Eismann, et al. A single negative charge within the pore region of a cGMP-gated channel controls rectification, Ca2+ blockage, and ionic selectivity. Proc. Natl Acad Sci U S A. Feb. 1, 1994;91(3):1109-13.

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