Modulation by bicarbonate, phosphate, and maleate of the kinetics of adenosinetriphosphatase activity and of the binding of manganese ions to chloroplast coupling factor 1.
Bicarbonate, maleate, and phosphate were shown to modulate adenosinetriphosphatase (ATPase) activity in coupling factor 1 from chloroplasts. Kinetic analysis of the changes in the ratio between the apparent Km with and without effectors indicated that the stimulation of the activity by bicarbonate was a result of a decrease in the Km for MnATP2-. The inhibition by phosphate resulted from a decrease in the Ki for free ATP as a competitive inhibitor at pH 8. THe effectors did not change Vmax at this pH. However, at pH 6.5, both Km and Vmax of ATPase activity with MnATP2- were changed by maleate, yet the mode of inhibition by free ATP remained unaltered. In addition to decreasing the Km, bicarbonate induced a 10-fold decrease in the Kd for binding of Mn2+ at the two tight binding sites in the presence of ATP at pH 8. At pH 6.5, maleate also decreased both the Km for MnATP2- and the Kd for Mn2+ binding. A decrease in the Km of a substrate induced by an effector is likely to be a result of a decrease in the binding constant of the substrate. Therefore, these results are in harmony with the suggested assignment of the two tight binding sites of Mn2+ at the active sites of the enzyme.
D000244:Adenosine Diphosphate; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate; D001639:Bicarbonates / Q000494:pharmacology*; D007700:Kinetics; D008298:Maleates / Q000494:pharmacology*; D008345:Manganese / Q000494:pharmacology*; D008433:Mathematics; D010710:Phosphates / Q000494:pharmacology*; D010944:Plants / Q000201:enzymology*; D011485:Protein Binding; D006180:Proton-Translocating ATPases / Q000378:metabolism*
D001639:Bicarbonates; D008298:Maleates; D010710:Phosphates; D008345:Manganese; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; C030272:maleic acid; D000251:Adenosine Triphosphatases; D006180:Proton-Translocating ATPases