Retrovirus-mediated expression of an artificial β-endorphin precursor in primary fibroblasts

A. S. Beutler, M. S. Banck, F. W. Bach, F. H. Gage, Frank Porreca, E. J. Bilsky, T. L. Yaksh

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Peptides are of potential interest in the field of gene therapy but require modification by genetic engineering to facilitate their secretion. Amino terminal addition of a signal peptide is not always sufficient to achieve this goal, as found in this study for β-endorphin. To overcome this problem, addition of the pre-pro-sequence of mouse nerve growth factor to β- endorphin was tested. Retrovirus-mediated expression of a hybrid construct of the pre-pro-sequence of nerve growth factor and human β-endorphin in primary fibroblasts resulted in the secretion of β-endorphin immunoreactivity at a rate of 620 pg/h/106 cells. Analysis of the secreted β-endorphin immunoreactivity with reverse-phase HPLC, immunoassays using three different antibodies, and an assay for the specific displacement of [3H][D-Ala2,N- MePhe4,Gly-ol5]enkephalin from μ-opioid receptors suggests that the pre- pro-sequence is cleaved off from the pre-pro-sequence/β-endorphin construct prior to secretion, resulting in bona fide β-endorphin. Transplantation of β-endorphin-secreting cells into brain or spinal cord may provide a gene therapy approach for the treatment of chronic, opioid-sensitive pain states.

Original languageEnglish (US)
Pages (from-to)475-481
Number of pages7
JournalJournal of Neurochemistry
Volume64
Issue number2
StatePublished - 1995

Fingerprint

Endorphins
Retroviridae
Fibroblasts
Gene therapy
Opioid Receptors
Nerve Growth Factor
Genetic Therapy
Genetic engineering
Genetic Engineering
Enkephalins
Protein Sorting Signals
Immunoassay
Opioid Analgesics
Assays
Brain
Spinal Cord
Transplantation
High Pressure Liquid Chromatography
Pain
Peptides

Keywords

  • Furin
  • Gene therapy
  • Nerve growth factor
  • Opioid peptides
  • Pain
  • Pre-pro-sequence

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Beutler, A. S., Banck, M. S., Bach, F. W., Gage, F. H., Porreca, F., Bilsky, E. J., & Yaksh, T. L. (1995). Retrovirus-mediated expression of an artificial β-endorphin precursor in primary fibroblasts. Journal of Neurochemistry, 64(2), 475-481.

Retrovirus-mediated expression of an artificial β-endorphin precursor in primary fibroblasts. / Beutler, A. S.; Banck, M. S.; Bach, F. W.; Gage, F. H.; Porreca, Frank; Bilsky, E. J.; Yaksh, T. L.

In: Journal of Neurochemistry, Vol. 64, No. 2, 1995, p. 475-481.

Research output: Contribution to journalArticle

Beutler, AS, Banck, MS, Bach, FW, Gage, FH, Porreca, F, Bilsky, EJ & Yaksh, TL 1995, 'Retrovirus-mediated expression of an artificial β-endorphin precursor in primary fibroblasts', Journal of Neurochemistry, vol. 64, no. 2, pp. 475-481.
Beutler, A. S. ; Banck, M. S. ; Bach, F. W. ; Gage, F. H. ; Porreca, Frank ; Bilsky, E. J. ; Yaksh, T. L. / Retrovirus-mediated expression of an artificial β-endorphin precursor in primary fibroblasts. In: Journal of Neurochemistry. 1995 ; Vol. 64, No. 2. pp. 475-481.
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