title: Mytilus edulis hemolymph contains pro-opiomelanocortin: LPS and morphine stimulate differential processing.
abstract: Mytilus edulis hemolymph contains mammalian-like proopiomelanocortin (POMC). The 20 kDa protein was purified by high pressure gel permeation chromatography, anti-adrenocorticotropin (ACTH)-affinity column and reverse-phase HPLC. The amino acid sequence determination was by Edman degradation, enzymatic treatments and Western blot analysis. Of the six peptides found in this opioid precursor, methionine-enkephalin, gamma-melanocyte stimulating hormone (MSH), alpha-MSH and ACTH exhibited 100, 80, 85 and 74% sequence identity, respectively, with the mammalian counterparts. beta-Endorphin and gamma-LPH exhibited only 25 and 10% sequence identity. Dibasic amino acid residues were found at the C-terminus of MSH and ACTH, indicating cleavage sites. The alpha-MSH is flanked at the C-terminus by Gly-Lys-Lys, representing an amidation signal. ACTH and CLIP (80% sequence identity) are also C-terminally flanked by dibasic amino acid residues. Furthermore, morphine, in a dose-dependent manner, increased the hemolymph levels of alpha-MSH and ACTH (1-39) in a naloxone and phosphoramidon antagonizable manner, indicating a neutral endopeptidase (24.11; NEP) mediated cleavage. Lipopolysaccharide (10 microg/animal) stimulated the processing of ACTH (1-39) yielding ACTH (1-24) in a cleavage that is independent of NEP, but dependent on aspartyl proteases, demonstrating differential enzymatic cleavage of ACTH (1-39). Taken together, POMC is present in invertebrates and its processing can be altered depending on the signal.
authors.affiliation: Multidisciplinary Center for the study of Aging, Neuroscience Research Institute, State University of New York, College at Old Westbury, Old Westbury, NY 11568-0210, USA.
authors.forename: G B
authors.initials: GB
authors.lastname: Stefano
authors.forename: B
authors.initials: B
authors.lastname: Salzet-Raveillon
authors.forename: M
authors.initials: M
authors.lastname: Salzet
mesh_terms: D000324:Adrenocorticotropic Hormone / Q000302:isolation & purification; D000595:Amino Acid Sequence; D000818:Animals; D049872:Bivalvia / Q000378:metabolism*; D017124:Conserved Sequence; D006458:Hemolymph / Q000187:drug effects* / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology*; D009074:Melanocyte-Stimulating Hormones / Q000378:metabolism*; D008969:Molecular Sequence Data; D009020:Morphine / Q000494:pharmacology*; D011333:Pro-Opiomelanocortin / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D017386:Sequence Homology, Amino Acid; D013268:Stimulation, Chemical
chemical_list: D008070:Lipopolysaccharides; D011333:Pro-Opiomelanocortin; D009020:Morphine; D000324:Adrenocorticotropic Hormone; D009074:Melanocyte-Stimulating Hormones