Effect of alkaline-phosphatase inhibition by 1- p-bromotetramisole on the formation of trichloroacetic acid-[ 32P]-insoluble phosphate from inorganic [ 32P]-phosphate and [ 32P]-pyrophosphate in non-mineralizing and mineralizing hamster molar tooth-germs in vitro

Abstract

In culture, 1- p-bromotetramisole ( pBTM), a specific inhibitor of alkaline phosphatase, significantly inhibited the formation of trichloroacetic acid (TCA)-insoluble [ 32P]-phosphate from inorganic [ 32P]-phosphate in the proliferating non-mineralizing second (M2) maxillary molar germs but had no effect in the actively mineralizing first (M1) germs. Addition of 10 −5 M inorganic pyrophosphate in the culture medium with a [ 32P]-phosphate label increased the inhibition of the formation of TCA-insoluble [ 32P]-phosphate in the M2. pBTM almost completely inhibited the formation of TCA-insoluble [ 32P]-phosphate from inorganic [ 32P]-pyrophosphate in the non-mineralizing M2. In the actively mineralizing M1, the compound significantly inhibited but did not abolish the formation of TCA-insoluble phosphate. These results confirm earlier biochemical findings that alkaline phosphatase possesses a pyrophosphatase activity probably related to the turnover of phosphorylated macromolecules necessary for cell differentiation and proliferation.