Neutral metalloendopeptidase associated with the smooth muscle cells of pregnant rat uterus.
The pregnant rat uterus contains a membrane-bound metalloendopeptidase that is biochemically and immunologically similar to kidney enkephalinase (E.C.3.4.24.11). The uterus enzyme readily cleaved specific neutral endopeptidase substrates and oxytocin as well as the synthetic elastase substrate, Suc(Ala)3-pNA, yet did not digest native elastin. Using specific inhibitors, the uterus endopeptidase was identified as a metallopeptidase and not a serine protease, having an absolute requirement for zinc and perhaps calcium for maximal activity. The uterus endopeptidase cross-reacted with polyclonal antiserum to kidney microvillar endopeptidase and a monoclonal antibody to common acute lymphocytic leukemia antigen. Immunohistochemical localization of the enzyme in a 17 day pregnant uterus indicated that the enzyme was localized on the smooth muscle bundles of the myometrium and the endometrial epithelium. Total enzyme activity was 25 times higher in the late-term pregnant uterus (17th day of pregnancy) than in the nonpregnant uterus. Enzyme levels dropped rapidly prior to parturition and within 4 days after delivery the enzyme activity had returned to control levels. Inhibition of NEP in uterine strips with phosphoramidon resulted in a marked potentiation of oxytocin-induced contractions. Our results suggest that the uterine endopeptidase may have an important role in regulating uterine smooth muscle cell contraction during the later stages of pregnancy through its action on oxytocin and perhaps other biologically active peptides.
Ottlecz
A
A
Department of Biochemistry, University of Texas Health Center, Tyler 75710.
Walker
S
S
Conrad
M
M
Starcher
B
B
D000818:Animals; D002458:Cell Fractionation; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D007700:Kinetics; D009130:Muscle, Smooth / Q000201:enzymology*; D015260:Neprilysin / Q000032:analysis / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010121:Oxytocin / Q000378:metabolism; D011247:Pregnancy; D051381:Rats; D013379:Substrate Specificity; D014590:Uterine Contraction / Q000502:physiology; D014599:Uterus / Q000201:enzymology*
D010121:Oxytocin; D015260:Neprilysin