Radiation inactivation analysis of H+-pyrophosphatase from submitochondrial particles of etiolated mung bean seedlings

Radiation inactivation analysis was employed to determine the functional masses of enzymatic activity and proton translocation of H+-pyrophosphatase from submitochondrial particles of etiolated mung bean seedlings. The activities of H+-pyrophosphatase decayed as a simple exponential function with respect to radiation dosage, D-37 values of 6.9+/-0.3 and 7.5 +/- 0.5 Mrad were obtained for pyrophosphate hydrolysis and its associated proton translocation, yielding molecular masses of 170+/-7 and 156+/-11 kDa, respectively. In the presence of valinomycin and 50 mM KCl, the functional size of H+-pyrophosphatase of tonoplast was decreased, while that of submitochondrial particles remained the same, indicating that they are two distinct types of proton pump using PPi as their energy source. (C) 2000 Federation of European Biochemical Societies.