Journal of Clinical Endocrinology & Metabolism, Vol 56, 1035-1041, Copyright © 1983 by Endocrine Society

ARTICLES

Adenosine 3',5'-monophosphate (cAMP)-binding protein and cAMP-dependent protein kinase in human placenta

JJ Moore, JV Baker and JA Whitsett

cAMP modulates estrogen, hCG, and lactate syntheses by human placenta, cAMP presumably exerts its major intracellular effect by binding to cAMP-dependent protein kinase (cAMP-PK), which, in turn, phosphorylates regulatory proteins within the target cell. cAMP binding and cAMP-PK have not been previously identified in placenta. [3H]cAMP binding to crude cytosol fractions of term placenta was rapid, saturable, and reversible. Scatchard analyses of saturation experiments of [3H]cAMP binding to placental cytosol were linear (Kd = 1.13 +/- 0.11 x 10(-8) M; n = 5). The binding capacity was 1.27 +/- 0.18 pmol/mg protein. Competition for the [3H]cAMP-binding site followed the potency order cAMP much greater than cGMP much greater than (Bu)2cAMP, analogous to cAMP binding to cAMP-PK in other tissues. ADP, ATP, and adenosine did not compete for the [3H]cAMP-binding site. cAMP significantly enhanced phosphorylation of histone protein by placental cytosol (activity ratio, 0.57 +/- 0.04; P less than 0.01). Two peaks of [3H]cAMP binding and coincident cAMP-PK activity were identified by DEAE-cellulose column chromatography of placental cytosol corresponding to classical type I and type II cAMP-PK. While the majority of the cAMP-PK was found in placental cytosol, cAMP-PK was also demonstrated in crude microsomal and microvillous brush border membranes of human placenta after solubilization with Triton X-100 (P less than 0.05). Regulation of placental function by catecholamines and other hormones known to mediate cAMP levels may be accomplished through the phosphorylation of cellular proteins by cAMP-dependent protein kinases.