Evidence for the presence of pro-oxytocin/neurophysin-converting enzyme in the human ovary.
The activity of pro-oxytocin/neurophysin (pro-OT/Np)-processing enzymes was determined in human granulosa cells, follicular fluids and purified secretory granules of corpora lutea. We detected the presence of an endoprotease which releases OT-Gly10-Lys11-Arg12 on cleavage of the synthetic pro-OT/Np(1-20) peptide after the dibasic Lys11-Arg12 doublet. This endoprotease was inhibited by EDTA, but was not affected by phenylmethanesulfonyl fluoride and pepstatin. Enzymatic activity was markedly reduced by replacement of L-Arg by D-Arg in the basic amino acid doublet of the substrate. The molecular weight of this enzyme was estimated to be 60 kDa. These features closely resembled those of the endoprotease identified in the bovine pituitary. The endoprotease is a metalloenzyme, specific for the Lys-Arg doublet. We also detected a carboxypeptidasic activity, inhibited by guanidinoethane-mercaptosuccinic acid. In the light of our previous detection of the processing intermediates, OT-Gly-Lys-Arg, OT-Gly-Lys and OT-Gly, in human ovary, these observations are in favour of a pro-OT/Np-processing pathway in the human ovary comparable with that in the bovine ovary. Moreover, these results confirm that oxytocin post-translational processing occurs in the human ovary and strongly suggest that pro-OT/Np is synthesized locally.
Guillou
M D
MD
Recherche Clinique et Biologique en Reproduction, INSERM CJF 89-08, Hôpital de la Grave, Toulouse, France.
Camier
M
M
Clamagirand
C
C
D002462:Cell Membrane / Q000201:enzymology; D002478:Cells, Cultured; D003338:Corpus Luteum / Q000201:enzymology / Q000648:ultrastructure; D003594:Cytoplasmic Granules / Q000201:enzymology; D010450:Endopeptidases / Q000032:analysis*; D005260:Female; D015571:Follicular Fluid / Q000201:enzymology; D006107:Granulosa Cells / Q000201:enzymology; D006801:Humans; D015592:Immunoradiometric Assay; D010053:Ovary / Q000201:enzymology*; D011863:Radioimmunoassay
D010450:Endopeptidases; C056535:pro-ocytocin-neurophysin convertase