Synenkephalin processing in embryonic rat brain.
Synenkephalin (proenkephalin 1-70) is produced and secreted as an intact molecule or as a part of precursors in the adult brain and adrenal medulla, respectively. However, it is cleaved to low molecular weight peptides in proliferating immune cells. Considering that the pre-proenkephalin gene is expressed in the embryonic rat brain during the cell proliferation stage, we studied the processing of synenkephalin in embryonic rat brains (E18) and compared it with the processing in adult rat brains. IR-synenkephalin was measured by RIA using a C-terminally directed antiserum. Adult rat brains contained higher concentrations of immunoreactive (IR)-synenkephalin (2,612 + 264) than embryonic rat brain (1,361 + 100) (results in fmol/mg proteins, n = 5). Gel filtration chromatography (Sephadex G-50) showed that in the extracts of adult rat brain, 50% of the IR-synenkephalin eluted in the position of the authentic peptide (8 kDa) and the rest of the immunoreactivity corresponded to partially processed peptides of 4.0 and 2.5 kDa. In embryonic rat brains synenkephalin was processed to intermediate peptides of 2.5, 1.7 and mainly to a low molecular weight peptide of 1.0 kDa. The concentration of this last peptide, which was further characterized by affinity column and HPLC, represented 45% of the total immunoreactivity. IR-met-enkephalin in embryonic rat brains (analyzed before and after enzymatic digestion with trypsin and carboxypeptidase B) corresponded principally to non-processed or partially processed products. However, these were cleaved to free met-enkephalin in adult rat brains.(ABSTRACT TRUNCATED AT 250 WORDS)
Rodriguez Vida
M I
MI
Instituto de Investigaciones Medicas, Seccion Sustancias Vasoactivas, Buenos Aires, Argentina.
Kleid
M C
MC
Ase
A
A
Finkielman
S
S
Nahmod
V E
VE
Vindrola
O
O
D000375:Aging / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001921:Brain / Q000166:cytology / Q000196:embryology*; D001923:Brain Chemistry / Q000502:physiology*; D002455:Cell Division; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004745:Enkephalins / Q000276:immunology / Q000378:metabolism*; D005260:Female; D017072:Neostriatum / Q000378:metabolism; D011247:Pregnancy; D011498:Protein Precursors / Q000276:immunology / Q000378:metabolism*; D011863:Radioimmunoassay; D051381:Rats; D017208:Rats, Wistar
D000911:Antibodies, Monoclonal; D004745:Enkephalins; D011498:Protein Precursors; C036693:synenkephalin