Plant DHDPR forms a dimer with unique secondary structure features that preclude higher-order assembly

Biochem J. 2018 Jan 5;475(1):137-150. doi: 10.1042/BCJ20170709.

Abstract

Dihydrodipicolinate reductase (DHDPR) catalyses the second reaction in the diaminopimelate pathway of lysine biosynthesis in bacteria and plants. In contrast with the tetrameric bacterial DHDPR enzymes, we show that DHDPR from Vitis vinifera (grape) and Selaginella moellendorffii are dimeric in solution. In the present study, we have also determined the crystal structures of DHDPR enzymes from the plants Arabidopsis thaliana and S. moellendorffii, which are the first dimeric DHDPR structures. The analysis of these models demonstrates that the dimer forms through the intra-strand interface, and that unique secondary features in the plant enzymes block tetramer assembly. In addition, we have also solved the structure of tetrameric DHDPR from the pathogenic bacteria Neisseria meningitidis Measuring the activity of plant DHDPR enzymes showed that they are much more prone to substrate inhibition than the bacterial enzymes, which appears to be a consequence of increased flexibility of the substrate-binding loop and higher affinity for the nucleotide substrate. This higher propensity to substrate inhibition may have consequences for ongoing efforts to increase lysine biosynthesis in plants.

Keywords: enzyme kinetics; lysine biosynthesis; oligomerisation; protein evolution.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs
  • Arabidopsis / chemistry
  • Arabidopsis / enzymology
  • Bacterial Proteins / chemistry*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism
  • Binding Sites
  • Coenzymes / chemistry
  • Coenzymes / metabolism
  • Crystallography, X-Ray
  • Dihydrodipicolinate Reductase / chemistry*
  • Dihydrodipicolinate Reductase / genetics
  • Dihydrodipicolinate Reductase / metabolism
  • Gene Expression
  • Kinetics
  • Lysine / biosynthesis
  • Models, Molecular
  • NAD / chemistry
  • NAD / metabolism
  • NADP / chemistry
  • NADP / metabolism
  • Neisseria meningitidis / chemistry
  • Neisseria meningitidis / enzymology
  • Picolinic Acids / chemistry*
  • Picolinic Acids / metabolism
  • Plant Proteins / chemistry*
  • Plant Proteins / genetics
  • Plant Proteins / metabolism
  • Protein Binding
  • Protein Conformation, alpha-Helical
  • Protein Conformation, beta-Strand
  • Protein Interaction Domains and Motifs
  • Protein Multimerization
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism
  • Selaginellaceae / chemistry
  • Selaginellaceae / enzymology
  • Species Specificity
  • Substrate Specificity
  • Vitis / chemistry
  • Vitis / enzymology*

Substances

  • 4-hydroxy-2,3,4,5-tetrahydro-2-dipicolinic acid
  • Bacterial Proteins
  • Coenzymes
  • Picolinic Acids
  • Plant Proteins
  • Recombinant Proteins
  • NAD
  • NADP
  • Dihydrodipicolinate Reductase
  • Lysine