title: The interaction of shikimic acid and protein phosphorylation with PEP carboxylase from the C4 dicot Amaranthus viridis.
abstract: Shikimic acid has been described as a potent competitive inhibitor of the activity of C4 phosphoenolpyruvate carboxylase (PEPC) from Amaranthus viridis. In the present study, the effects of shikimic acid were examined further with the dephospho (dark-form) and in vitro phosphorylated forms of homogeneous PEPC from A. viridis. Kinetic analysis showed that the inhibitor effect of shikimic acid was dependent on the phosphorylation state of the enzyme. Thus, the I50 value of shikimic acid for dark-form PEPC was six times lower than that for the phosphorylated enzyme (12 vs 71 microM, respectively). When Glc6P, an activator of C4 PEPC, was present in the assay medium, the I50 value increased 2- and 3-times with the phospho and dephospho PEPC-forms, respectively. Shikimic acid also markedly decreased 32P incorporation from Mg[gamma-32P]ATP into the dark-form of C4 PEPC, but not casein, catalyzed by protein kinase A. In this way, shikimic acid mimics the behaviour of L-malate, a well-known inhibitor of PEPC, in that it decreases both the enzyme's activity and phosphorylatability. Based on these data, a possible role for shikimic acid in the regulation of PEPC activity in plants is suggested.
authors.affiliation: Universidad Nacional de Rosario (UNR), Argentina.
authors.forename: S L
authors.initials: SL
authors.lastname: Colombo
authors.forename: C S
authors.initials: CS
authors.lastname: Andreo
authors.forename: R
authors.initials: R
authors.lastname: Chollet
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D003720:Densitometry; D004591:Electrophoresis, Polyacrylamide Gel; D007700:Kinetics; D010730:Phosphoenolpyruvate Carboxylase / Q000378:metabolism*; D010766:Phosphorylation; D018515:Plant Leaves / Q000201:enzymology*; D012765:Shikimic Acid / Q000378:metabolism*
chemical_list: D012765:Shikimic Acid; D000255:Adenosine Triphosphate; D010730:Phosphoenolpyruvate Carboxylase