Reconstitution of the purified gamma-aminobutyric acid-benzodiazepine receptor complex from bovine cerebral cortex into phospholipid vesicles.
The purified gamma-aminobutyric acid-benzodiazepine receptor complex from bovine cerebral cortex has been reconstituted into phospholipid vesicles by a cholate dialysis procedure. The reconstituted receptor bound [3H]flunitrazepam at a single class of sites with dissociation constant Kd = 2.9 +/- 0.3 nM, an increase in affinity to the membrane level from the 4-fold weakening found in detergent solution. It also bound [3H]muscimol with a Kd for the high-affinity sites of approximately 50 nM. [35S]tert.-Butyl-bicyclophosphorothionate, for which there is evidence in membranes for binding to a channel gating site on this receptor, showed similar binding to the reconstituted receptor.
D000818:Animals; D002417:Cattle; D002540:Cerebral Cortex / Q000032:analysis*; D002784:Cholesterol; D008081:Liposomes; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; D010743:Phospholipids; D011963:Receptors, GABA-A / Q000302:isolation & purification
D008081:Liposomes; D010713:Phosphatidylcholines; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; D010743:Phospholipids; D011963:Receptors, GABA-A; C024990:asolectin; D002784:Cholesterol