ATP-ase activity in the odontoblastic layer of rat incisor Determination with a radiochemical and a colorimetric method

Abstract

The ATP-splitting enzyme activity in odontoblasts isolated from rat incisors has been studied by means of a radiochemical and a colorimetric micromethod. The results with the two methods were virtually identical. The reaction was linear with time for at least 45 min. The pH optimum was found to be 9.8 independently of the ATP concentration. Maximal substrate saturation occurred at a total ATP concentration of 3 mM. Ca²⁺ and Mg²⁺ ions activated ATP degradation. F- ions did not affect the activity at low concentrations, whereas higher concentrations were inhibitory. Na⁺ and K⁺ ions had no influence on ATP splitting enzyme activity, while PO4^3- ions were slightly inhibitory. Urea inhibited the enzyme activity at concentrations above 1.5 M, while EDTA and EGTA were strong inhibitors at very low concentrations. When incubating in the presence of low concentrations of specific inhibitors for nonspecific alkaline phosphatase, levamisole and R 8231, about 20% ATP degrading enzyme activity remained. In conclusion it is suggested that there are at least two ATP degrading phosphatases active at alkaline pH.

Key Words:

• Biochemistry
• mineralization
• dentinogenesis