In:
ChemBioChem, Wiley, Vol. 7, No. 9 ( 2006-09-04), p. 1400-1409
Abstract:
The quantification of pharmacological parameters at G protein‐coupled receptors (GPCRs) is indispensable in drug research but costly and time‐consuming when conventional methods are sequentially applied. With neuropeptide Y (NPY) Y 1 , Y 2 , and Y 5 receptors as model systems, a homogenous flow cytometric method for the simultaneous determination of the affinity, selectivity, and activity of GPCR ligands was developed. Mixtures of cells expressing the receptors of interest and cyanine‐labeled NPY as a universal fluorescent Y 1 , Y 2 , and Y 5 receptor agonist were used. Calcium mobilization was measured in different channels with the aid of fluo‐4 and fura red. A combination of dye‐loaded HEL‐Y 1 and CHO‐Y 2 ‐Gα qi5 cells with unloaded HEC‐1B‐Y 5 cells allowed the simultaneous determination of Y 1 , Y 2 , and Y 5 receptor selectivity preceded by the Y 1 and Y 2 receptor‐mediated response with one and the same sample. The data are in good agreement with those determined by radioligand binding and spectrofluorimetry. The convenient, robust, and inexpensive multiparametric procedure offers a broad range of applications in the pharmacological characterization of GPCR ligands.
Type of Medium:
Online Resource
ISSN:
1439-4227
,
1439-7633
DOI:
10.1002/cbic.200600163
Language:
English
Publisher:
Wiley
Publication Date:
2006
detail.hit.zdb_id:
2020469-3
SSG:
12
Bookmarklink