Isolation and characterization of an endogenous C-terminal fragment of the alpha-neo-endorphin/dynorphin precursor from bovine caudate nucleus.
Antibodies have been raised to a synthetic peptide corresponding to the C-terminal 15-amino acid residues of prodynorphin, the common precursor to the neo-endorphins and dynorphins. The amino acid sequence of the antigen was based on the sequence deduced from mRNA isolated and cloned from porcine hypothalamus (Kakidani, H., Y. Furutani, H. Takahashi, M. Noda, Y. Morimoto, T. Hirose, M. Asai, S. Inayama, S. Nakanishi, and S. Numa (1982) Nature 298: 245-248). Using a radioimmunoassay developed from these antibodies we have isolated an endogenous prodynorphin C-fragment from bovine caudate nucleus. The isolated peptide displayed characteristics on gel filtration similar to those of synthetic prodynorphin C-fragment predicted from the porcine mRNA sequence but had low cross-reactivity in the radioimmunoassay. Sequencing and amino acid analysis showed a substitution of serine for asparagine at position 6 in the porcine sequence. Dynorphin B (rimorphin), which is adjacent to prodynorphin C-fragment in the precursor, was isolated from the same extract. Amino acid analysis and elution position on a gel filtration column confirmed its structure as that previously characterized from bovine pituitary extracts. The release of prodynorphin C-fragment and the C-terminus of dynorphin B from the porcine precursor would require cleavage at a single arginine residue. However, a terminal arginine was not present on either of these prodynorphin peptides isolated from bovine caudate. The data would suggest that processing at a single arginine residue results in elimination of the arginine, a feature in common with processing at paired basic residues.
Evans
C J
CJ
Barchas
J D
JD
Esch
F S
FS
Böhlen
P
P
Weber
E
E
D000818:Animals; D002417:Cattle; D002421:Caudate Nucleus / Q000032:analysis*; D004399:Dynorphins / Q000031:analogs & derivatives* / Q000032:analysis; D004723:Endorphins / Q000032:analysis*; D004745:Enkephalins / Q000032:analysis*; D011498:Protein Precursors / Q000032:analysis*; D011817:Rabbits; D011863:Radioimmunoassay
D004723:Endorphins; D004745:Enkephalins; D011498:Protein Precursors; D004399:Dynorphins; C036200:rimorphin; C033660:preproenkephalin