Molecular aspects of the regulation of rat kidney alkaline phosphatase.
The mechanisms by which phosphate regulates the activity of alkaline phosphatase (orthophosphoric monoester phosphohydrolase, EC 3.1.3.1) in rat kidney were investigated. Measurements of incorporation of [(14)C]leucine into kidney alkaline phosphatase in rats fed on complete or phosphate-free diet provide evidence of a twofold increase in the rate of synthesis of the enzyme in diet-treated animals. Cycloheximide experiments indicated that control and diet-adapted enzyme decreases in activity according to first-order kinetics with a calculated half-life of 10.3 and 6.5h after complete and phosphate-free diet administration respectively. Basal and diet-adapted enzymes exhibit similar K(m) values for several phosphomonoesters and an identical degree of inhibition is produced by cysteine. In addition, the enzyme from both sources is the same with regard to heat inactivation at 45, 56 or 64 degrees C, to the profile of elution from Sephadex and to electrophoretic properties on polyacrylamide gel. A failure of rat kidney alkaline phosphatase to respond to cortisol (hydrocortisone) was also observed.
D000311:Adrenal Glands / Q000502:physiology; D000315:Adrenalectomy; D000469:Alkaline Phosphatase / Q000032:analysis / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000378:metabolism*; D000818:Animals; D002247:Carbon Isotopes; D002850:Chromatography, Gel; D003513:Cycloheximide / Q000494:pharmacology; D003545:Cysteine / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D004032:Diet; D004589:Electrophoresis, Disc; D006207:Half-Life; D006358:Hot Temperature; D006854:Hydrocortisone / Q000494:pharmacology; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D007930:Leucine / Q000378:metabolism; D008297:Male; D010710:Phosphates / Q000378:metabolism*; D051381:Rats
D002247:Carbon Isotopes; D010710:Phosphates; D003609:Dactinomycin; D003513:Cycloheximide; D000469:Alkaline Phosphatase; D007930:Leucine; D003545:Cysteine; D006854:Hydrocortisone