Effects of Medium Viscosity Increasing Agents on ATP Synthesis in Chloroplast Thylakoids
The effect of an increase in the medium viscosity on cyclic photophosphorylation in chloroplast thylakoids and on Ca2+-dependent ATP hydrolysis by the chloroplast coupling factor CF1 was studied. With 0.1-0.2 mM ADP used it was found that the rate of ATP synthesis decreases after addition of various agents that increase the medium viscosity (sucrose, dextran 40 or polyethylene glycol 6000 provided that these agents cause neither uncoupling nor electron transport inhibition in the absence of ADP. Dextran and polyethylene glycol inhibited ATP synthesis by 50% when their concentrations were much lower (6-10%) than that of sucrose (30-40%), while 50% inhibition of Ca2+-dependent ATP hydrolysis by CF1-ATPase was observed at higher concentrations of dextran and polyethylene glycol (9-13 %) and lower concentrations of sucrose (about 20%). For ADP, the effective Michaelis constant (KM) was shown to increase 2-3-fold with the increasing viscosity; meanwhile the maximal rate of cyclic photophosphorylation remained virtually unchanged. The dependence of KМ on the medium viscosity can serve as a criterion for the process of diffusioncontrolled photophosphorylation. Possible mechanisms of ADP and ATP diffusion are discussed.