Evolution of C(4) phosphoenolpyruvate carboxylase in Flaveria: determinants for high tolerance towards the inhibitor L-malate.
During the evolution of angiosperms, C4 phosphoenolpyruvate carboxylases have evolved several times independently from ancestral non-photosynthetic isoforms. They show distinct kinetic and regulatory properties when compared with the C3 isozymes. To identify the evolutionary alterations which are responsible for C4-specific properties, particularly the increased tolerance towards the allosteric inhibitor L-malate, the photosynthetic phosphoenolpyruvate carboxylase of Flaveria trinervia Mohr C4 and its ortholog from the closely related C3 plant Flaveria pringlei Gand. were examined using reciprocal enzyme chimeras. The main determinants for a high tolerance towards L-malate were located in the C-terminal region of the C4 enzyme. The effect of interchanging the region between amino acids 296 and 437 was strongly dependent upon the activation of the enzyme by glucose-6-phosphate. This confirms earlier observations that this region is important for the regulation of the enzyme by glucose-6-phosphate and that it harbours determinants for the different response of the C3 and the C4 enzyme towards this allosteric activator. In addition, it was possible to demonstrate that the only C4-specific amino acid, a serine in the C-terminal part of the enzyme, is not involved in conferring an increased L-malate tolerance to the C4 enzyme.
Jacobs
Bianca
B
Institut für Entwicklungs-und Molekularbiologie der Pflanzen, Heinrich-Heine Universität Düsseldorf, Universitätsstrasse 1, D-40225 Düsseldorf, Germany.
Engelmann
Sascha
S
Westhoff
Peter
P
Gowik
Udo
U
D000595:Amino Acid Sequence; D004791:Enzyme Inhibitors; D019143:Evolution, Molecular; D037142:Flaveria / Q000201:enzymology* / Q000235:genetics*; D018506:Gene Expression Regulation, Plant / Q000502:physiology; D019298:Glucose-6-Phosphate; D008293:Malates / Q000494:pharmacology*; D008969:Molecular Sequence Data; D019764:Phosphoenolpyruvate Carboxykinase (ATP) / Q000037:antagonists & inhibitors* / Q000235:genetics* / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism
D004791:Enzyme Inhibitors; D008293:Malates; D010940:Plant Proteins; D019298:Glucose-6-Phosphate; C030298:malic acid; D019764:Phosphoenolpyruvate Carboxykinase (ATP)