A novel opioid mechanism seems to modulate phagocytosis in Tetrahymena.
We have previously reported that a beta-endorphin-like substance inhibits phagocytosis in Tetrahymena perhaps by a mu-like opioid receptor. We now report a further characterization of the elements involved in the signal transduction mechanism of this opioid. Affinity chromatography followed by immunoblots of both intracellular extracts and extracellular medium reveal the presence of two main proteins of 64 and 75 kDa. These molecular weights are much higher than that of any known opioid peptide or precursor protein and suggest that we may be dealing with either a novel opioid or with proteins that by chance cross-react with anti-beta-endorphin antibody. Nevertheless, when the biological activity of these proteins was tested it was found that they had an effect similar to that of mammalian beta-endorphin, namely inhibition of phagocytosis by a naloxone-reversible mechanism. We have probed a size-selected Tetrahymena library with a pro-opiomelanocortin probe and have obtained several positive clones; the sequencing of their inserts should establish whether we are dealing with a bona fide member of the opioid family. Another aspect we have been studying is the G-proteins which appear to be involved in the modulation of phagocytosis. We have found, by means of Western blotting (using an antibody against the conserved GTP-binding region of the alpha-subunit), two bands of 51 and 59 kDa; no alpha-subunit of 59 kDa had been reported previously and may represent a novel G-protein. In spite of these differences, the opioid signal transduction mechanism appears to remarkably resemble that present in more complex organisms.
D000818:Animals; D015139:Blotting, Southern; D003429:Cross Reactions; D004271:DNA, Fungal / Q000032:analysis; D015151:Immunoblotting; D008322:Mammals; D009270:Naloxone / Q000494:pharmacology; D010587:Phagocytosis / Q000187:drug effects; D011333:Pro-Opiomelanocortin / Q000096:biosynthesis; D051381:Rats; D011957:Receptors, Opioid / Q000502:physiology; D013768:Tetrahymena / Q000502:physiology*; D001615:beta-Endorphin / Q000302:isolation & purification / Q000494:pharmacology / Q000502:physiology*
D004271:DNA, Fungal; D011957:Receptors, Opioid; D009270:Naloxone; D001615:beta-Endorphin; D011333:Pro-Opiomelanocortin