According to the membrane compartment concept the receptor specificity of ligands is based not only on ligand-receptor complementarity but also on specific ligand-membrane interactions. Elaboration of this concept for opioid peptide-receptor interactions had led to the assumption that mu- and delta-receptors are located in anionic and cationic membrane compartments, respectively, and to the prediction that positively charged opioid receptor ligands should display mu-receptor selectivity. To assess the validity of this model, we synthesized a series of dermorphin analogues carrying a net positive charge and tested them in mu- and delta-receptor representative binding assays and bioassays. Some but not all of the prepared compounds showed the receptor-selectivity profile expected on the basis of the membrane compartment concept. In particular, gradual augmentation of the positive charge from 1+ to 3+ in a series of dermorphin-(1-4) tetrapeptide analogues produced an enhancement of mu-receptor affinity and a progressive decrease in delta-receptor affinity, resulting in increasingly higher mu-receptor selectivity. The most selective compound was [D-Arg2,Lys4]dermorphin-(1-4)-amide (DALDA), showing a selectivity ratio (Ki delta/Ki mu = 11,400) more than 10 times higher than that of DAGO (Ki delta /Ki mu = 1050) and, thus, displaying unprecedented mu-receptor specificity. Because of its high positive charge (3+), DALDA may be particularly useful as a very specific agonist for studying peripheral mu-receptor interactions.