Fusicoccin- and light-induced activation and in vivo phosphorylation of phosphoenolpyruvate carboxylase in vicia guard cell protoplasts.

The in vivo regulation of phosphoenolpyruvate carboxylase (PEPCase; EC 4.1.1.31), was studied in purified guard cell protoplasts (GCPs) of Vicia faba L. Incubation of GCPs with fusicoccin (FC) led to the rapid activation of PEPCase and reduced its sensitivity towards the feedback-inhibitor malate. This was accompanied by an increase in the phosphorylation state of the enzyme. Additionally, PEPCase could be transiently activated by white light. Activation and phosphorylation of PEPCase upon illumination were dependent on the presence of potassium in the incubation medium. Treatment of GCPs with inhibitors of H(+)-ATPases, and with abscisic acid (ABA) suppressed the activation of PEPCase in a concentration-dependent manner. Treatment of protoplasts with butyrate also led to PEPCase activation, suggesting a role for the cytosolic pH (pH(cyt)) in the signal transduction process. The presented data indicate that guard cell PEPCase is regulated by reversible phosphorylation of at least one isoform and elucidate first components of the signaling pathway.