Synthesis, opiate receptor affinity, and conformational parameters of [4-tryptophan]enkephalin analogues

A series of analogues of the opioid peptide enkephalin with tryptophan substituted for phenylalanine in position 4 was synthesized by the solid-phase method. The [Trp4]enkephalin analogues and the corresponding [Phe4]enkephalin analogues displayed nearly parallel affinities in the opiate receptor binding assay throughout the series. In a conformational study fluorescence parameters were measured and intramolecular Tyr-Trp distances were estimated on the basis of resonance energy transfer experiments. No gross conformational differences were observed between analogues with widely differing opiate receptor affinity; however, small but significant changes in the intramolecular distance between the phenol ring and the indole moiety and/or in their relative orientation became apparent in some compounds. Identical intramolecular distances of 9.3 +/- 0.2 angstrom between the two aromatic rings were obtained with [Trp4,Met5]enkephalin, [Trp4,Leu5]enkephalin, and the N-terminal tetrapeptide comprised in the latter two analogues, indicating the existence of folded conformationas in 2 X 10(-5) M aqueous solution and demonstrating conformational analogy between these three peptides. The conformational parameters are discussed in relation to the observed affinities and the putative opiate receptor topography.