Further characterisation of the [35S]-TBPS binding site of the GABA receptor complex in locust (Schistocerca gregaria) ganglia membranes.
1. Using homogenates of supraoesophageal ganglia from locust we observed specific binding of [35S]-TBPS which was linear with protein concentration up to 7 mg/ml, showed a pH optimum at pH 9.0 and was linear with NaCl concentration up to 350 mM. 2. Kinetic analysis of the binding showed positive cooperativity as a result of changes in on and off-rates with occupation of the binding site by the ligand. The apparent KD = 417 nM and Bmax = 1083 fmol/mg of membrane protein were calculated using a computer program for dose-response curve fitting. 3. The binding was enhanced by GABA, pentobarbital and benzodiazepines. Picrotoxinin had no effect on the binding at 0.1 mM. Only the cage convulsants TBPS and IBP were able to displace the binding. 4. Whilst the characteristics of the binding are similar to those reported for house fly thorax and abdomen preparations they are significantly different from those reported for house fly head, cockroach nerve cord and rat brain.
Brown
M C
MC
Department of Biochemistry, University of Bath, UK.
Lunt
G C
GC
Stapleton
A
A
D000818:Animals; D001569:Benzodiazepines / Q000494:pharmacology; D005724:Ganglia / Q000032:analysis*; D006110:Grasshoppers / Q000032:analysis*; D008024:Ligands; D008566:Membranes / Q000032:analysis; D010424:Pentobarbital / Q000494:pharmacology; D011963:Receptors, GABA-A / Q000032:analysis* / Q000187:drug effects; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology
D008024:Ligands; D011963:Receptors, GABA-A; C041598:t-butylbicyclophosphorothionate receptor; D001569:Benzodiazepines; D005680:gamma-Aminobutyric Acid; D010424:Pentobarbital