Characterization of the F1F0-ATPase and the tightly-bound ATPase activities in submitochondrial particles from human term placenta.

In a previous study we demonstrated the existence of a tightly-bound ATPase in the human placental mitochondria (Martínez et al., 1993). The current study characterizes the ATP hydrolysis produced by the F1F0-ATPase and the tightly-bound ATPase in submitochondrial particles from the human term placenta. Both enzymes were not differentiated by pH. Inhibitors were necessary to distinguish the activity of each enzyme. The kinetic of the total ATP hydrolysis fitted into a model of two enzymes. During the characterization, it was observed that the tightly-bound ATPase activity was partially inhibited by vanadate and Mg2+, whereas the F1F0-ATPase was totally inhibited by Mg2+. Different nucleotides were hydrolyzed by the tightly-bound ATPase; the F1F0-ATPase hydrolyzed exclusively ATP. Glucose-6-phosphate, p-nitrophenylphosphate, or pyrophosphate were not hydrolyzed by the F1F0-ATPase, although some hydrolysis was observed with the tightly-bound ATPase. It is concluded that the tightly-bound ATPase activity corresponded to a 5'-nucelotidase, and that the human placental mitochondria could participate in the metabolism of nucleotides.