The effects of magnesium on nucleoside phosphatase activity in frog skin

Abstract

Histochemical tests, employing the Wachstein-Meisel medium, indicate that nucleoside triphosphatase activity is found predominantly in two areas of the frog skin epidermis: (1) in mitochondria, where activity is enhanced by dinitrophenol, Mg(2+) dependent, but inhibited by fixation; and (2) apparently associated with cell membranes of the middle and outer portions of the epidermis, where activity is inhibited by Mg(2+), unaffected by dinitrophenol, and only slightly reduced by fixation. Spectrophotometric analysis shows that Mg(2+) in the medium does not increase spontaneous hydrolysis of ATP, thus obviating the possible explanation that changes in substrate concentrations in the medium lead to alterations in the "staining" distributions. It is postulated that perhaps the two enzymes differ in their requirements for substrate-one requiring the polyphosphate to be in complexed form with Mg(2+), the other uncomplexed. Concentrations of Mg(2+) required to inhibit cell membrane nucleoside triphosphatase activity also inhibit the electrical potential difference and short-circuit current of the frog skin. Although these observations might be taken as presumptive evidence of the cell membrane enzyme as a component of the ion pump system, because of certain dissimilarities with respect to the biochemists' "transport ATPase" and for other reasons discussed in the paper, any definite conclusions in this regard are premature.