Characterization and expression of uterine and placental alkaline phosphatases in the mouse.
Alkaline phosphatase (ALP) is rapidly induced in the uterine subepithelial stroma after a natural or artificial decidual stimulus. During gestation ALP-specific activity peaked at Day 7 to 8 (Day 1 is day of detection of the copulation plug) followed by a rapid decline to control levels by Day 9. This elevation in enzyme activity was preceded by an 8-fold induction of a 2.6 kilobase (kb) mRNA. This mRNA was not preferentially localized to implantation sites. ALP activity was detected in the placenta at Day 9 and reached maximum specific activity at Day 19. The placental ALP was also encoded by a 2.6 kb mRNA. Uterine and placental ALPs were inhibited to the same extent by levamisole, L-tryptophan and homoarginine. The calculated Ki values for these inhibitors were not statistically different between the uterine and placental forms. Km values towards the substrate p-nitrophenylphosphate, however, were statistically different between the uterine and placental forms. Both uterine and placental ALPs were stimulated 3-4-fold by addition of 2 mM-Mg2+. Electrophoretic mobilities on SDS polyacrylamide gel, where the enzyme migrated as a single band, were the same. The uterine form, however, could be distinguished from the placental isoenzyme by separation on non-denaturing polyacrylamide gels; the uterine form had a single zone of activity which migrated with an intermediate mobility between the two zones of activity detected for the placental enzyme. These differences in mobility could be ascribed to the sialic acid content of the enzyme because treatment with neuraminidase resulted in the uterine and placental forms migrating with comparable but slower mobilities in non-denaturing gels.(ABSTRACT TRUNCATED AT 250 WORDS)
Pollard
J W
JW
Department of Developmental Biology and Cancer, Albert Einstein College of Medicine, Bronx, NY 10461.
Jahan
M
M
Butterworth
P J
PJ
D000469:Alkaline Phosphatase / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D015152:Blotting, Northern; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006709:Homoarginine / Q000494:pharmacology; D007978:Levamisole / Q000494:pharmacology; D051379:Mice; D010052:Ovariectomy; D010920:Placenta / Q000187:drug effects / Q000201:enzymology*; D011247:Pregnancy; D012333:RNA, Messenger / Q000032:analysis; D014364:Tryptophan / Q000494:pharmacology; D014599:Uterus / Q000187:drug effects / Q000201:enzymology*
D012333:RNA, Messenger; D006709:Homoarginine; D007978:Levamisole; D014364:Tryptophan; D000469:Alkaline Phosphatase