TY - JOUR
T1 - Processing of prodynorphin in BRL-3A cells, a rat liver-derived cell line
T2 - implications for the specificity of neuropeptide-processing enzymes
AU - Petanceska, Suzana
AU - Zikherman, Julie
AU - Pricker, Lloyd D.
AU - Devi, Lakshmi
N1 - Funding Information:
This work was supportedi n part by NINDS grant number NS-26880( to L.D) and NIDA grant number DA-04494 (to L.D.F.). We would like thank Leanne Daly for technicala ssistance.
PY - 1993/7
Y1 - 1993/7
N2 - 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.
AB - 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.
KW - Carboxypeptidase E
KW - Carboxypeptidase H
KW - Dynorphin
KW - Dynorphin converting enzyme
KW - Furin
KW - Insulin-like growth factor
KW - Neuropeptide
KW - Prohormone convertase
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U2 - 10.1016/0303-7207(93)90049-P
DO - 10.1016/0303-7207(93)90049-P
M3 - Article
C2 - 8375574
AN - SCOPUS:0027200452
SN - 0303-7207
VL - 94
SP - 37
EP - 45
JO - Molecular and Cellular Endocrinology
JF - Molecular and Cellular Endocrinology
IS - 1
ER -