Characterization of flunitrazepam and beta-carboline high affinity binding in bovine pineal gland
High affinity binding of3H-fiunitrazepam (FNZP) to crude membrane preparations of bovine pineal membranes was examined by a rapid filtration procedure through Whatman GFB paper. At 0 °C binding reached equilibrium in about 20 min. Scatchard analysis of data at equilibrium revealed a single populatio...
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todo:paper_00283835_v37_n2_p150_Lowenstein2023-10-03T14:38:49Z Characterization of flunitrazepam and beta-carboline high affinity binding in bovine pineal gland Lowenstein, P.R. Cardinali, D.P. Barbiturates Benzodiazepine binding GABA Pineal gland β-Carboline binding 4 aminobutyric acid 4' chlorodiazepam barbituric acid derivative benzodiazepine clonazepam flumazenil flunitrazepam pentobarbital radioisotope animal cell beta carboline 3 carboxylic acid ethyl ester h 3 cattle cell membrane central nervous system drug receptor binding endocrine system flunitrazepam h 3 nonhuman pharmacokinetics pineal body Animal Anti-Anxiety Agents, Benzodiazepine Benzodiazepines Binding Sites Carbolines Cattle Comparative Study Flunitrazepam gamma-Aminobutyric Acid In Vitro Indoles Kinetics Pentobarbital Pineal Gland Rats Species Specificity Support, Non-U.S. Gov't High affinity binding of3H-fiunitrazepam (FNZP) to crude membrane preparations of bovine pineal membranes was examined by a rapid filtration procedure through Whatman GFB paper. At 0 °C binding reached equilibrium in about 20 min. Scatchard analysis of data at equilibrium revealed a single population of binding sites with dissociation constant (K<inf>d</inf>) = 3.14 ± 0.45 nM and binding site concentration (B<inf>max</inf>) = 55.6 ± 5.6 fmol/mg protein. Kinetic analysis of the association and dissociation curves indicated a kinetic K<inf>d</inf> = 1.13 nM, in reasonable agreement to that obtained at equilibrium. When various benzodiazepine (BZP) analogues were tested for their ability to inhibit3H-FNZP binding, the following K<inf>i</inf>(nM) were obtained: Clonazepam (0.22), Ro 15-1788(0.48), FNZP (0.95), Ro 5-4864 (> 10,000). When the β-carboline derivative3H-ethyl-β-carboline-3-carboxylate ester (EβCEE) was used as a radioligand, K<inf>d</inf> at equilibrium (0.98 nM), kinetic K<inf>d</inf> (1.69 n M) and affinity order for analogues were in close agreement to those found for FNZP binding; however, B<inf>max</inf> was about 60% that observed for3H-FNZP binding. Addition of GABA or pentobarbital (100 μM) to pineal membranes increased3H-FNZP binding by 55 and 72%. These results suggest the existence of a mixed population of type 1 and type 2 central BZP receptor subclass in bovine pineal gland. © 1983 S. Karger AG, Basel. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00283835_v37_n2_p150_Lowenstein |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Barbiturates Benzodiazepine binding GABA Pineal gland β-Carboline binding 4 aminobutyric acid 4' chlorodiazepam barbituric acid derivative benzodiazepine clonazepam flumazenil flunitrazepam pentobarbital radioisotope animal cell beta carboline 3 carboxylic acid ethyl ester h 3 cattle cell membrane central nervous system drug receptor binding endocrine system flunitrazepam h 3 nonhuman pharmacokinetics pineal body Animal Anti-Anxiety Agents, Benzodiazepine Benzodiazepines Binding Sites Carbolines Cattle Comparative Study Flunitrazepam gamma-Aminobutyric Acid In Vitro Indoles Kinetics Pentobarbital Pineal Gland Rats Species Specificity Support, Non-U.S. Gov't |
| spellingShingle |
Barbiturates Benzodiazepine binding GABA Pineal gland β-Carboline binding 4 aminobutyric acid 4' chlorodiazepam barbituric acid derivative benzodiazepine clonazepam flumazenil flunitrazepam pentobarbital radioisotope animal cell beta carboline 3 carboxylic acid ethyl ester h 3 cattle cell membrane central nervous system drug receptor binding endocrine system flunitrazepam h 3 nonhuman pharmacokinetics pineal body Animal Anti-Anxiety Agents, Benzodiazepine Benzodiazepines Binding Sites Carbolines Cattle Comparative Study Flunitrazepam gamma-Aminobutyric Acid In Vitro Indoles Kinetics Pentobarbital Pineal Gland Rats Species Specificity Support, Non-U.S. Gov't Lowenstein, P.R. Cardinali, D.P. Characterization of flunitrazepam and beta-carboline high affinity binding in bovine pineal gland |
| topic_facet |
Barbiturates Benzodiazepine binding GABA Pineal gland β-Carboline binding 4 aminobutyric acid 4' chlorodiazepam barbituric acid derivative benzodiazepine clonazepam flumazenil flunitrazepam pentobarbital radioisotope animal cell beta carboline 3 carboxylic acid ethyl ester h 3 cattle cell membrane central nervous system drug receptor binding endocrine system flunitrazepam h 3 nonhuman pharmacokinetics pineal body Animal Anti-Anxiety Agents, Benzodiazepine Benzodiazepines Binding Sites Carbolines Cattle Comparative Study Flunitrazepam gamma-Aminobutyric Acid In Vitro Indoles Kinetics Pentobarbital Pineal Gland Rats Species Specificity Support, Non-U.S. Gov't |
| description |
High affinity binding of3H-fiunitrazepam (FNZP) to crude membrane preparations of bovine pineal membranes was examined by a rapid filtration procedure through Whatman GFB paper. At 0 °C binding reached equilibrium in about 20 min. Scatchard analysis of data at equilibrium revealed a single population of binding sites with dissociation constant (K<inf>d</inf>) = 3.14 ± 0.45 nM and binding site concentration (B<inf>max</inf>) = 55.6 ± 5.6 fmol/mg protein. Kinetic analysis of the association and dissociation curves indicated a kinetic K<inf>d</inf> = 1.13 nM, in reasonable agreement to that obtained at equilibrium. When various benzodiazepine (BZP) analogues were tested for their ability to inhibit3H-FNZP binding, the following K<inf>i</inf>(nM) were obtained: Clonazepam (0.22), Ro 15-1788(0.48), FNZP (0.95), Ro 5-4864 (> 10,000). When the β-carboline derivative3H-ethyl-β-carboline-3-carboxylate ester (EβCEE) was used as a radioligand, K<inf>d</inf> at equilibrium (0.98 nM), kinetic K<inf>d</inf> (1.69 n M) and affinity order for analogues were in close agreement to those found for FNZP binding; however, B<inf>max</inf> was about 60% that observed for3H-FNZP binding. Addition of GABA or pentobarbital (100 μM) to pineal membranes increased3H-FNZP binding by 55 and 72%. These results suggest the existence of a mixed population of type 1 and type 2 central BZP receptor subclass in bovine pineal gland. © 1983 S. Karger AG, Basel. |
| format |
JOUR |
| author |
Lowenstein, P.R. Cardinali, D.P. |
| author_facet |
Lowenstein, P.R. Cardinali, D.P. |
| author_sort |
Lowenstein, P.R. |
| title |
Characterization of flunitrazepam and beta-carboline high affinity binding in bovine pineal gland |
| title_short |
Characterization of flunitrazepam and beta-carboline high affinity binding in bovine pineal gland |
| title_full |
Characterization of flunitrazepam and beta-carboline high affinity binding in bovine pineal gland |
| title_fullStr |
Characterization of flunitrazepam and beta-carboline high affinity binding in bovine pineal gland |
| title_full_unstemmed |
Characterization of flunitrazepam and beta-carboline high affinity binding in bovine pineal gland |
| title_sort |
characterization of flunitrazepam and beta-carboline high affinity binding in bovine pineal gland |
| url |
http://hdl.handle.net/20.500.12110/paper_00283835_v37_n2_p150_Lowenstein |
| work_keys_str_mv |
AT lowensteinpr characterizationofflunitrazepamandbetacarbolinehighaffinitybindinginbovinepinealgland AT cardinalidp characterizationofflunitrazepamandbetacarbolinehighaffinitybindinginbovinepinealgland |
| _version_ |
1807324176518742016 |