Abstract
1. The activity of pyruvate,P(i) dikinase in leaves of maize and Amaranthus palmeri rapidly falls on transferring illuminated plants to darkness. Illumination of dark-treated plants results in an immediate rapid increase in activity of the enzyme, the final activity reached being dependent on the intensity of the incident light. 2. Activation of the enzyme in extracts of dark-treated maize leaves after gel filtration on Sephadex G-25 requires a thiol and P(i). The P(i) requirement for activation can be replaced by arsenate. Activation of the enzyme is inhibited by AMP and GMP and possibly also by ADP and ATP. Activation of the enzyme after gel filtration on Sephadex G-200 also requires a heat-labile component that is excluded by Sephadex G-25. 3. The active enzyme isolated from illuminated leaves is inactivated by ADP in the presence of a thiol, the rate of inactivation being very much faster in air than in an oxygen-free atmosphere. Reactivation of the ADP-inactivated enzyme requires a thiol, P(i) and a component excluded by Sephadex G-25 but considerably retarded by Sephadex G-200. 4. The active enzyme is rapidly and irreversibly inactivated in the absence of a thiol. Inactivation is accelerated by both sodium diethyldithiocarbamate and tetraethylthiuram disulphide, and the enzyme inactivated by these reagents is completely reactivated by incubation with dithiothreitol. This reactivation does not require P(i). The inactive enzyme from dark-treated leaves is stabilized by diethyldithiocarbamate and can be partially activated by dithiothreitol alone; complete reactivation requires both dithiothreitol and P(i). 5. The enzyme activity is markedly inhibited by the thiol reagents p-chloromercuribenzoate, gamma-(p-arsenophenyl)-n-butyrate and an equimolar mixture of arsenite and 2,3-dimercaptopropan-1-ol. 6. The processes of activation and inactivation observed in vitro are discussed in relation to the regulation of pyruvate,P(i) dikinase activity in the leaf.