Ionic strength and pH effects on the rates of reduction of spinach plastocyanin by ascorbate

Kinetic studies of the reduction of plastocyanin from spinach chloroplasts by ascorbate were carried out using a stopped flow apparatus. The reduction of spinach plastocyanin by ascorbate followed first-order kinetics in the reductant with a rate constant k = 415 M-1.S-1 [20 degrees C, 0.1 M NaCl, pH 8.0 (10 mM Tris-HCl)]. The second-order rate constants for the reduction of plastocyanin by ascorbate increased dramatically with increasing pH and ionic strength. The cationic micelles of cetyltrimethylammonium bromide also accelerated this electron-transfer reaction. The importance of electrostatic interactions between plastocyanin and ascorbate is discussed in relation to the photosynthetic electron-transport reactions.