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Functional screening of adrenergic receptors by measuring intracellular calcium using the FlexStation scanning fluorimeter
Author(s) -
Nanda Kamna,
Chatterjee Mou,
Gupta Suman,
Singh Rakesh,
Tiwar Atul,
Gupta Dikshi,
Ray Abhijit
Publication year - 2009
Publication title -
biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.144
H-Index - 84
eISSN - 1860-7314
pISSN - 1860-6768
DOI - 10.1002/biot.200800205
Subject(s) - radioligand , g protein coupled receptor , receptor , radioligand assay , adrenergic receptor , intracellular , chemistry , recombinant dna , terazosin , affinities , calcium in biology , second messenger system , pharmacology , biology , biochemistry , endocrinology , hyperplasia , gene
In this study we test whether functional screening of compounds to adrenergic G protein‐coupled receptors (GPCRs) would provide data that correlated significantly with radiolabeled binding data, thereby permitting researchers to replace expensive radioligand‐binding experiments with non‐radioactive screening. An increase in intracellular calcium levels represents an important second messenger signal for several recombinant GPCRs. In this study, we describe the affinities of three alpha adrenoceptor antagonists (terazosin, tamsulosin and alfuzosin), determined by monitoring the changes in intracellular calcium levels and comparing them with their radioligand‐binding affinities. In addition to determining the functional affinities of the three alpha adrenoceptor antagonists, we evaluate their binding at two alpha adrenoceptor subtypes and optimized the assay for high‐throughput screening.