Opioid receptor affinity of multivalent ligand system consisting of polymerized liposome

A multivalent ligand system, in which enkephalin/phospholipid conjugates were immobilized on a polymerized liposome, has been prepared. A hydrophilic spacer chain, -(Sar-Sar-Pro)2-, was placed between Tyr-D-Ala-Gly-Trp-Leu (an enkephalin part) and a phospholipid part to diminish steric hindrance of the liposome against receptor binding of the enkephalin unit. The affinity of the immobilized enkephalin conjugate for opioid receptor was dependent on the density of the enkephalin unit on the surface of polymerized liposome, indicating that more than two enkephalin units on a liposome simultaneously bind to receptors in the membrane. IC50 values for the delta- and mu-receptors were 15 and 26 nM, respectively, when the conjugate was immobilized at the molar ratio of [enkephalin]/[phospholipid] of 340. The latter affinity is better than that of Tyr-D-Ala-Gly-Trp-Leu (50 nM). The enkephalin part was located at the membrane surface, as indicated by fluorescence spectroscopy. When 12% cerebroside sulfate was mixed in the polymerized liposome, delta-receptor affinity of the immobilized enkephalin conjugate was improved to 7.4 nM. The environment around the enkephalin part became hydrophilic upon incorporation of cerebroside sulfate into the polymerized liposome, resulting in changes in the receptor affinities.