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Characterization of benzodiazepine receptors with fluorescent ligands
Author(s) -
McCabe R. Tyler,
Gosta Brian R.,
Miller Rachel L.,
Havunjian R. Hratchia,
Rige Kenner C.,
Skolnick Phil
Publication year - 1990
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.4.11.2165950
Subject(s) - fluorescence , benzodiazepine , receptor , chemistry , gabaa receptor , biophysics , biochemistry , biology , physics , quantum mechanics
Fluorescein conjugates of the high‐affinity benzodiazepine receptor ligands Ro 15‐1788 and Ro 7‐1986 were synthesized. The binding of these fluorescent ligands (BD 621 and BD 607) to benzodiazepine receptors was characterized by direct fluorescence measurement. Both the equilibrium dissociation constants ( K D ) of BD 621 and BD 607 and the maximum number of binding sites ( B max ) estimated by fluorescence monitoring were consistent with values obtained by using radioligand binding techniques. The binding of BD 621 and BD 607 assessed by fluorescence measurement was reversible, abolished by photoaffinity labeling with Ro 15‐4513, and unaffected by a variety of substances that do not bind to benzodiazepine receptors. The potencies of chemically diverse benzodiazepine receptor compounds to inhibit fluorescent ligand binding were highly correlated ( r = 0.94, P < 0.001), with potencies obtained from radioligand binding techniques. These findings demonstrate the feasibility of using direct fluorescence measurement techniques to quantitate ligand‐receptor interactions.— M c C abe , R. T.; de C osta , B. R.; M iller , R. L.; H avunjian , R. H.; R ice , K. C.; S kolnick , P. Characterization of benzodiazepine receptors with fluorescent ligands. FASEB J. 4: 2934‐2940; 1990.