Prodynorphin in invertebrates.
We have characterized a prodynorphin-like molecule in an invertebrate, specifically in the rhynchobdellid leech Theromyzon tessulatum. The 14270 Da protein was purified by gel permeation chromatography, anti-leucine-enkephalin-affinity column separation followed by reverse-phase HPLC. Its complete characterization was performed by Edman degradation, enzymatic treatments, and matrix assisted laser-desorption time of flight mass spectrometry. This 119 amino-acid protein exhibits 28.8% sequence identity with rat prodynorphin, 22.9% with human prodynorphin, and 21.8% with the pig molecule. Within the leech precursor, alpha-Neo-endorphin, dynorphin-A, and dynorphin B-like peptides are present at the C-terminus as in vertebrate prodynorphin. These biological active peptides exhibit 100%, 50%, and 76.6% sequence identity with their counterparts in mammals, respectively. The amount of leucine-enkephalin is identical to that found in vertebrates. Leech prodynorphin is distinguished from that found in mammals in that the N-terminus is shorter. This report constitutes the first complete biochemical characterization of a prodynorphin in invertebrates.