[3H]Prazosin bound to a single class of high-affinity sites in both bovine aortic and rat hepatic membranes. The absolute affinity values of displacing ligands (prazosin greater than doxazosin much greater than trimazosin greater than yohimbine) were the same for both tissues. After solubilization of the alpha 1-adrenoceptors with digitonin and 3-[(3-cholamidopropyl)-dimethylammonio]-1-propane sulfonate, an identical rank order potency was observed. However, solubilization significantly reduced ligand affinity. In both tissues the affinity of prazosin was reduced 10- to 13-fold, whereas the affinities of doxazosin, trimazosin and yohimbine were reduced two- to six-fold. There appeared to be no relationship between the lipophilicities of the ligands and the degree to which affinity is affected by solubilization. The results suggest that the reductions in affinity are the consequence of a conformational change in the alpha 1-adrenoceptor and appear to support the hypothesis that the alpha 1-adrenoceptor is so constructed that the spatial configuration of the binding site can change in response to an alteration in its microenvironment.