Production of bioactive enkephalin from the nonendocrine cell lines COS-7, NIH3T3, Ltk-, and C2C12.
Enkephalin is synthesized from proenkephalin in neuroendocrine cells. For the attempt to induce nonneuroendocrine origin cells to produce enkephalin, we used a mammalian expression vector for fusion peptides, pMEproCT beta, in which a fused peptide is designed to be cleaved by a yeast Kex2-like endoprotease furin. Met-Enkephalin was expressed in four nonneuroendocrine cell lines: COS-7, C2C12, Ltk-, and NIH3T3. The four cell lines produced a marked amount of Met-enkephalin, which appeared as a single peak on reverse-phase HPLC. Because transplantation of adrenal medullary cells to the subarachnoid space has been used to alleviate terminal cancer pain, and enkephalin appears to play a central role in relieving pain, this enkephalin expression vector may be useful for direct enkephalin expression in pericancerous tissues.
D016475:3T3 Cells; D000199:Actins / Q000235:genetics; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D043423:Carboxypeptidase H; D002268:Carboxypeptidases / Q000378:metabolism; D002460:Cell Line; D002645:Chickens; D002522:Chlorocebus aethiops; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D000242:Cyclic AMP / Q000378:metabolism; D004744:Enkephalin, Methionine / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D045683:Furin; D005822:Genetic Vectors; D007739:L Cells; D008322:Mammals; D051379:Mice; D008969:Molecular Sequence Data; D009270:Naloxone / Q000494:pharmacology; D009838:Oligodeoxyribonucleotides; D010957:Plasmids; D011401:Promoter Regions, Genetic; D043484:Proprotein Convertases; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000494:pharmacology; D015183:Restriction Mapping; D012441:Saccharomyces cerevisiae / Q000201:enzymology; D029701:Saccharomyces cerevisiae Proteins; D013381:Subtilisins / Q000378:metabolism*; D014162:Transfection; D014407:Tumor Cells, Cultured
D000199:Actins; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D009270:Naloxone; D004744:Enkephalin, Methionine; D000242:Cyclic AMP; D002268:Carboxypeptidases; D043423:Carboxypeptidase H; D043484:Proprotein Convertases; D013381:Subtilisins; C058751:KEX2 protein, S cerevisiae; D045683:Furin