Reconstitution of α1-adrenoceptors having high affinity for prazosin

Abstract Using a variety of detergents we find that soluble α 1 -adrenoceptor recovery from aortic and hepatic membranes is markedly enhanced if the receptor is first prelabelled with prazosin. Moreover, prelabelling prevents the reduction in prazosin affinity induced by solubilizing concentrations of digitonin, possibly by stabilizing the receptor's conformation, 20–25% of α 1 -adrenoceptors solubilized in sodium cholate were reconstituted into brain lipids. Specific [ 3 H]prazosin binding to the reconstituted receptor was saturable and of high affinity (K D = 0.019 ± 0.008 nM). We conclude that prelabelling is essential to preserve the receptor in detergents. However, once the detergent is removed brain lipids alone are able to maintain the receptor in a form with high affinity for prazosin.