PMID: 995Sep 1, 1975

The mechanism of hydrolysis of beta-glycerophosphate by kidney alkaline phosphatase

The Biochemical Journal
J Ahlers

Abstract

1. To identify the functional groups that are involved in the conversion of beta-glycerophosphate by alkaline phosphatase (EC 3.1.3.1) from pig kidney, the kinetics of alkaline phosphatase were investigated in the pH range 6.6-10.3 at substrate concentrations of 3 muM-30 mM. From the plots of log VH+ against pH and log VH+/KH+m against pH one functional group with pK = 7.0 and two functional groups with pK = 9.1 were identified. These groups are involved in substrate binding. Another group with pK = 8.8 was found, which in its unprotonated form catalyses substrate conversion. 2. GSH inhibits the alkaline phosphatase reversibly and non-competitively by attacking the bound Zn(II). 3. The influence of the H+ concentration on the activation by Mg2+ ions of alkaline pig kidney phosphate was investigated between pH 8.4 and 10.0. The binding of substrate and activating Mg2+ ions occurs independently at all pH values between 8.4 and 10.0. The activation mechanism is not affected by the H+ concentration. The Mg2+ ions are bound by a functional group with a pK of 10.15. 4. A scheme is proposed for the reaction between enzyme, substrate, Mg2+ and H+ and the overall rate equation is derived. 5. The mechanism of substrate binding and splitt...Continue Reading

Citations

Oct 1, 1992·European Journal of Biochemistry·T C ChangG G Chang
Jul 1, 1986·Annals of Human Genetics·D E WilsonS Povey
May 23, 2015·Journal of Environmental Science and Health. Part. B, Pesticides, Food Contaminants, and Agricultural Wastes·Ahmed F El-Aswad, Mohamed E I Badawy
Jul 1, 1983·Cell Biochemistry and Function·P J Butterworth
Jul 1, 1983·Cell Biochemistry and Function·D W Howat, J Chayen
Dec 1, 1978·Annals of the Rheumatic Diseases·J J JohnstoneJ Chayen
Nov 24, 1976·Calcified Tissue Research·R SheddenJ Chayen
Nov 1, 1985·Naunyn-Schmiedeberg's Archives of Pharmacology·K H Leser, H Osswald
May 20, 2015·Macromolecular Bioscience·Werner E G MüllerXiaohong Wang
Jul 6, 2017·Journal of Applied Oral Science : Revista FOB·Jia Tang, Takashi Saito

Related Concepts

Alkaline Phosphatase
Reduced Glutathione
Calcium Glycerophosphate
Hydrogen-Ion Concentration
Hydrolysis
Kidney
Magnesium
Zinc

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