Processing of prodynorphin in BRL-3A cells, a rat liver-derived cell line

implications for the specificity of neuropeptide-processing enzymes

Suzana Petanceska, Julie Zikherman, Lloyd D. Fricker, Lakshmi Devi

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Prodynorphin is post-translationally processed into dynorphin B-13 and other peptides by the action of endopeptidases that cleave at pairs of basic amino acids and at single basic residues, followed by a Carboxypeptidase that removes the C-terminal basic residues. To evaluate the specificity of neuropeptide processing enzymes, rat prodynorphin was transfected into BRL-3A cells, a rat liver-derived cell line which produces insulin-like growth factor II, but does not normally express prodynorphin. The transfected prodynorphin was post-translationally processed at both monobasic and dibasic cleavage sites, with the formation of dynorphin B-13 and other peptides. This finding indicates that BRL-3A cells express prodynorphin-processing enzymes. These cells were found to secrete two enzyme activities previously implicated in the processing of dynorphin, a monobasic cleaving 'dynorphin converting enzyme' and 'carboxypeptidase E', based on inhibitor sensitivities and pH optima. The dynorphin converting enzyme secreted from BRL-3A cells elutes from an anion exchange column under the same conditions as the enzyme secreted from pituitary-derived cell lines (AtT-20, GH4C1). Northern blot analysis indicates that BRL-3A cells express Carboxypeptidase E mRNA in addition to mRNA encoding furin, a prohormone-processing endopeptidase. The mRNAs for two other related endopeptidases, prohormone convertase 1 and 2, were not detected on Northern blots, suggesting that these enzymes are not required for the processing of prodynorphin. The expression of Carboxypeptidase E, furin, and dynorphin converting enzyme in BRL-3A cells suggests that these peptide processing enzymes are not specific for neuropeptides, but are also present in cells which process peptide growth factors.

Original languageEnglish (US)
Pages (from-to)37-45
Number of pages9
JournalMolecular and Cellular Endocrinology
Volume94
Issue number1
DOIs
StatePublished - 1993

Fingerprint

Neuropeptides
Liver
Rats
Carboxypeptidase H
Cells
Cell Line
Endopeptidases
Enzymes
Processing
Furin
Peptides
Messenger RNA
Northern Blotting
Proprotein Convertase 2
Proprotein Convertase 1
Carboxypeptidases
Dynorphins
Basic Amino Acids
Cathepsin A
Insulin-Like Growth Factor II

Keywords

  • Carboxypeptidase E
  • Carboxypeptidase H
  • Dynorphin
  • Dynorphin converting enzyme
  • Furin
  • Insulin-like growth factor
  • Neuropeptide
  • Prohormone convertase

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Processing of prodynorphin in BRL-3A cells, a rat liver-derived cell line : implications for the specificity of neuropeptide-processing enzymes. / Petanceska, Suzana; Zikherman, Julie; Fricker, Lloyd D.; Devi, Lakshmi.

In: Molecular and Cellular Endocrinology, Vol. 94, No. 1, 1993, p. 37-45.

Research output: Contribution to journalArticle

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