Photosynthesis Research

, 88:73

The regulatory role of residues 226–232 in phosphoenolpyruvate carboxylase from maize

Authors

  • Jiping Yuan
    • Department of Chemistry & BiochemistryCalifornia State University
  • Joyce Sayegh
    • Department of Chemistry & BiochemistryCalifornia State University
  • Julian Mendez
    • Department of Chemistry & BiochemistryCalifornia State University
  • Laurell Sward
    • Department of Chemistry & BiochemistryCalifornia State University
  • Norma Sanchez
    • Department of Chemistry & BiochemistryCalifornia State University
  • Susan Sanchez
    • Department of Chemistry & BiochemistryCalifornia State University
  • Grover Waldrop
    • Department of BiochemistryLouisiana State University
    • Department of Chemistry & BiochemistryCalifornia State University
Regular paper

DOI: 10.1007/s11120-005-9032-x

Cite this article as:
Yuan, J., Sayegh, J., Mendez, J. et al. Photosynth Res (2006) 88: 73. doi:10.1007/s11120-005-9032-x

Abstract

The regulatory properties of maize phosphoenolpyruvate carboxylase were significantly altered by site-directed mutagenesis of residues 226 through 232. This conserved sequence element, RTDEIRR, is part of a surface loop at the dimer interface. Mutation of individual residues in this sequence caused various kinetic changes, including desensitization of the enzyme to key allosteric effectors or alteration of the K0.5 PEP for the substrate phosphoenolpyruvate. R231A, and especially R232Q, displayed decreased apparent affinity for the activator glucose-6-phosphate. Apparent affinity for the activator glycine was reduced in D228N and R232Q, while the maximum activation caused by glycine was greatly reduced in R226Q and E229A. R226Q and E229A also showed significantly lower sensitivity to the inhibitors malate and aspartate. E229A exhibited a low K0.5 PEP, while the K0.5 PEP of R232Q was significantly higher than that of wild type. Thus these seven residues are critical determinants of the enzyme’s kinetic responses to activators, inhibitors and substrate. The present results support an earlier suggestion that Arg 231 contributes to the binding site of the allosteric activator glucose-6-phosphate, and are consistent with other proposals that the substrate phosphoenolpyruvate allosterically activates the enzyme by binding at or near the glucose-6-phosphate site. The results also suggest that the glycine binding site may be contiguous with the glucose-6-phosphate binding site. Glu 229, which extends from this interface region through the interior of the protein and emerges near the aspartate binding site, may provide a physical link for propagating conformational changes between the allosteric activator and inhibitor binding regions.

Keywords

allosteric regulationC4 photosynthesisglucose-6-phosphate activationglycine activationmalate inhibitionphosphoenolpyruvate carboxylasesite-directed mutagenesis

Abbreviations

CAM

Crassulacean Acid Metabolism

G6P

glucose-6-phosphate

PEP

phosphoenolpyruvate

Copyright information

© Springer Science+Business Media, Inc. 2006