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, pMEproCTβ, in which a fused peptide is designed to be cleaved by a yeast Kex2-like endoprot...

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Bibliographic Details
Published in:Peptides (New York, N.Y. : 1980) N.Y. : 1980), 1995, Vol.16 (5), p.933-938
Main Authors: Takahashi, Ken-Ichiro, Fujita, Tatsushi, Takeuchi, Toshiyuki
Format: Article
Language:English
Subjects:
3T3 Cells
Actins - genetics
Amino Acid Sequence
Animals
Base Sequence
C2C12
Carboxypeptidase H
Carboxypeptidases - metabolism
Cell Line
Cercopithecus aethiops
Chickens
Chromatography, High Pressure Liquid - methods
COS-7
Cyclic AMP - metabolism
Enkephalin
Enkephalin, Methionine - biosynthesis
Enkephalin, Methionine - isolation & purification
Enkephalin, Methionine - pharmacology
Furin
Fusion peptide
Genetic Vectors
L Cells (Cell Line)
Ltk
Mammals
Mice
Molecular Sequence Data
Naloxone - pharmacology
NIH3T3
Oligodeoxyribonucleotides
Plasmids
Processing
Promoter Regions, Genetic
Proprotein Convertases
Protein Processing, Post-Translational
Recombinant Fusion Proteins - biosynthesis
Recombinant Proteins - biosynthesis
Recombinant Proteins - isolation & purification
Recombinant Proteins - pharmacology
Restriction Mapping
Saccharomyces cerevisiae - enzymology
Saccharomyces cerevisiae Proteins
Subtilisins - metabolism
Transfection
Tumor Cells, Cultured
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Summary: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, pMEproCTβ, 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.
ISSN:0196-9781
1873-5169
DOI:10.1016/0196-9781(95)00061-N