title: Alkaline phosphatase (EC 3.1.3.1) in serum is inhibited by physiological concentrations of inorganic phosphate.
abstract: Natural and artificial manipulation of tissue nonspecific alkaline phosphatase activity indicates that pyrophosphate, phosphoethanolamine, and pyridoxal 5'-phosphate are among the natural substrates for this enzyme. Although inorganic phosphate has been recognized as a competitive inhibitor of this enzyme for many years, the influence of phosphate on alkaline phosphatase activity in serum under physiological conditions has not been previously reported. We examined the kinetics of tissue nonspecific alkaline phosphatase from bovine kidney and sera from 49 patients with a wide range of endogenous phosphate concentrations using pyridoxine 5'-phosphate as a substrate at pH 7.4. For the bovine kidney enzyme, the Km was 0.42 +/- 0.04 micromol/L, and the Ki for phosphate was 2.4 +/- 0.2 micromol/L. Analysis of the kinetics using pyridoxine 5'-phosphate in undiluted serum from 10 subjects with phosphorus ranging from 0.5-2.1 mmol/L and alkaline phosphatase activity ranging from 41-165 nmol/min x mL gave estimates for the Km of 56 +/- 11 micromol/L and for the Ki of 540 +/- 82 micromol/L for phosphate. This indicates that under physiological conditions alkaline phosphatase activity toward pyridoxine 5'-phosphate is reduced approximately 50% by the normal phosphate concentration and that it will increase or decrease significantly in response to changes in phosphate concentration within the ranges observed clinically.
authors.affiliation: Biochemistry Department, Fort Wayne State Developmental Center, Indiana 46835, USA. coburn@ipfw.edu
authors.forename: S P
authors.initials: SP
authors.lastname: Coburn
authors.forename: J D
authors.initials: JD
authors.lastname: Mahuren
authors.forename: M
authors.initials: M
authors.lastname: Jain
authors.forename: Y
authors.initials: Y
authors.lastname: Zubovic
authors.forename: J
authors.initials: J
authors.lastname: Wortsman
mesh_terms: D000328:Adult; D000368:Aged; D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors / Q000097:blood*; D000818:Animals; D002417:Cattle; D004791:Enzyme Inhibitors / Q000097:blood*; D005260:Female; D006801:Humans; D007422:Intestines / Q000201:enzymology; D007668:Kidney / Q000201:enzymology; D008297:Male; D008875:Middle Aged; D009928:Organ Specificity; D010710:Phosphates / Q000097:blood*; D010920:Placenta / Q000201:enzymology; D013379:Substrate Specificity
chemical_list: D004791:Enzyme Inhibitors; D010710:Phosphates; D000469:Alkaline Phosphatase