Adenylate cyclase from term human placenta and its regulation

L. Milewich, S. Hendricks, J. E. Graham, N. F. Gant, B. E. Schwarz, P. C. MacDonald

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

The activation of placental AC by either Mg2+ or Mn2+, in the presence and absence of NaF, followed sigmoidal saturation kinetics. Mn2+ enhanced maximally the NaF-stimulated Mg2+-dependent AC activity. The apparent Km of Mg2+-dependent AC for ATP was 0.4 mM, with and without NaF addition. GTP and GMP-P(NH)P stimulated the Mg2+-dependent AC in a dose-dependent manner with half-maximal stimulation taking place at concentrations of approximately 2 μM. In the presence of GMP-P(NH)P (10 μm) the kinetics of the AC dependence on Mg+ ion concentration changed from sigmoidal to hyperbolic. Most of the AC activity (> 83 per cent) was associated with the particulate fractions of placental homogenate. For better reproducibility, the AC assay was performed using sonicated particulate fraction preparations; sonication did not alter the response of AC to stimuli to a variety of agents used in these experiments; freezing and thawing, however, obliterated the stimulation by β-adrenergic agonists. Placental AC activity was inhibited by p-hydroxymercuriphenyl sodium sulphonate in a dose-dependent fashion, and the inhibition was reversed by dithiothreitol. Mg2+-dependent AC was inhibited by 0.5 mm phenylhydrazine (95 per cent). Mg2+-dependent AC activity was responsive to stimulation by epinephrine, without and with GTP addition, with half-maximal stimulation taking place at a concentration of 2 μm. The stimulatory effect of epinephrine was blocked by propranolol in a dose-dependent manner but was not blocked by phentolamine. Oestrone, oestradiol-17β, 2-hydroxyoestreone, 2-hydroxyoestradiol-17β, dehydroepiandrosterone sulphate, and progesterone, as well as oxytocin, did not alter either the basal or GMP-P(NH)P-stimulated Mg2+-dependent AC activities. Preincubation of 20 000 g particulate fraction with either NaF or GMP-P(NH)P, followed by washing, resulted in preparations that remained stimulated without the requirement of any further additions.

Original languageEnglish (US)
Pages (from-to)165-180
Number of pages16
JournalPlacenta
Volume3
Issue number2
DOIs
StatePublished - Jan 1 1982

ASJC Scopus subject areas

  • Reproductive Medicine
  • Obstetrics and Gynecology
  • Developmental Biology

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    Milewich, L., Hendricks, S., Graham, J. E., Gant, N. F., Schwarz, B. E., & MacDonald, P. C. (1982). Adenylate cyclase from term human placenta and its regulation. Placenta, 3(2), 165-180. https://doi.org/10.1016/S0143-4004(82)80050-7