Adenosine 3',5'-monophosphate-dependent phosphoproteins in human placenta.
cAMP modulates estrogen, hCG, and lactate syntheses by the human placenta. The major effects of cAMP are presumably mediated through the phosphorylation of specific regulatory phosphoproteins after cAMP activation of cAMP-dependent protein kinase. cAMP-dependent phosphoproteins have not been identified in the placenta. Homogenates and cytosol from term human placenta were subjected to an endogenous protein phosphorylation assay using [gamma-32P]ATP in the presence and absence of 1.0 microM cAMP. Protein phosphorylation was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. cAMP stimulated 32P incorporation into proteins with mol wt of 25,000, 27,000, 39,000, 45,000, 52,000, 58,000, and 73,000 (P less than 0.02). Half-maximal 32P incorporation was observed with 1.0 X 10(-7) M cAMP, which was similar to the concentration required for half-maximal histone kinase activity (8.5 +/- 2.9 X 10(-8) M). cGMP induced 32P incorporation into the same placental proteins as cAMP, but 2 orders of magnitude greater cGMP concentrations were required to achieve phosphorylation levels similar to those caused by cAMP. cAMP-dependent protein kinase inhibitor completely blocked cGMP-induced phosphorylation, even when histone protein was added. Therefore, no evidence of a cGMP-dependent protein kinase or specific cGMP-dependent phosphoproteins were detected. CaCl2 (10(-8) - 10(-4) M) had no effect on cAMP-induced 32P incorporation into the seven cAMP-dependent phosphoproteins. However calcium induced 32P incorporation into four other proteins (mol wt, 97,000, 90,000, 20,000, and 19,000). Regulation of placental metabolism by catecholamines and other hormones known to mediate intracellular cAMP or calcium levels may be accomplished by phosphorylation of these phosphoproteins.
Moore
J J
JJ
Cardaman
R C
RC
D002118:Calcium / Q000494:pharmacology; D000242:Cyclic AMP / Q000494:pharmacology*; D006152:Cyclic GMP / Q000494:pharmacology; D003600:Cytosol / Q000378:metabolism; D005260:Female; D006801:Humans; D008274:Magnesium / Q000494:pharmacology; D008970:Molecular Weight; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D010920:Placenta / Q000187:drug effects / Q000378:metabolism*; D011247:Pregnancy; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases / Q000378:metabolism
D010750:Phosphoproteins; D047428:Protein Kinase Inhibitors; D000242:Cyclic AMP; D011494:Protein Kinases; D006152:Cyclic GMP; D008274:Magnesium; D002118:Calcium