Evaluation of the interactions of mu and delta selective ligands with [ 3H]D-ala 2-D-Leu 5-enkephalin binding to mouse brain membranes

Abstract

The interactions of putative mu and delta selective ligands with [ 3H]D-ala 2-D-leu 5 enkephalin (DADLE) binding to mouse brain membranes were investigated. Computerized curve fitting of displacement curves performed at three different concentrations of 3H-DADLE indicated that a one site competitive model was sufficient to explain the interactions of leu-enkephalin (LE) and D-ser 2-thr 6-leucine enkephalin with 3H-DADLE binding. Similar experiments with morphine and morphiceptin were unique in that the multiple displacement curves crossed over one another. A two-site competitive model was required to adequately describe the interactions of these mu selective ligands with 3H-DADLE. This two-site model was one in which the inhibitor had higher affinity for the site labeled with lower affinity by 3H-DADLE. However, this two site model did not correctly predict the interaction of LE with 3H-DADLE in the presence of morphiceptin. These data indicate that: 1) putative mu and delta selective ligands do not bind to a common high affinity site; 2) mu selective ligands are not simple mixed inhibitors of a single site labeled by 3H-DADLE; and 3) competitive binding models may not explain the interaction of mu ligands with 3H-DADLE binding.