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Influence of δ‐Opioid Receptors on Production of Labeled Methionine 5 ‐Enkephalin in Murine Neuroblastoma Cells
Author(s) -
Gilbert Judith A.,
Richelson Elliott
Publication year - 1985
Publication title -
journal of neurochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.75
H-Index - 229
eISSN - 1471-4159
pISSN - 0022-3042
DOI - 10.1111/j.1471-4159.1985.tb12905.x
Subject(s) - methionine , pentapeptide repeat , enkephalin , receptor , intracellular , chemistry , biochemistry , clone (java method) , medicine , endocrinology , microbiology and biotechnology , biology , amino acid , opioid , peptide , dna
Synthesis of methionine 5 ‐enkephalin by intact cells of murine neuroblastoma clone N1E‐115 has been demonstrated both immunocytochemically and biochemically. In addition, N1E‐115 cells possess homogeneous enkephalin (δ) receptors which inhibit prostaglandin E 1 ‐ induced intracellular cyclic AMP formation. An assay was developed for measuring de novo synthesis of methionine 5 ‐enkephalin by pulsing cells in culture with radioactive methionine and isolating this pentapeptide to radiochemical purity by a procedure that included immunoaffinity chromatography specific for oxidized methionine 5 ‐enkephalin. This assay indicated that production of radiolabeled‐methionine 5 ‐enkephalin was increased upon lengthy exposure of intact N1E‐115 cells in the late logarithmic phase of growth to a nonproteolyz‐able analog of methionine 5 ‐enkephalin. This increase in synthesis of intracellular methionine 5 ‐enkephalin relative to control cells was prevented by prior incubation of the clone with naloxone, indicating that the response was mediated by the δ receptor.