Cloning, expression, purification and crystallization of dihydrodipicolinate synthase from Agrobacterium tumefaciens

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2012 Sep 1;68(Pt 9):1040-7. doi: 10.1107/S1744309112033052. Epub 2012 Aug 30.

Abstract

Dihydrodipicolinate synthase (DHDPS) catalyzes the first committed step of the lysine-biosynthesis pathway in bacteria, plants and some fungi. This study describes the cloning, expression, purification and crystallization of DHDPS (NP_354047.1) from the plant pathogen Agrobacterium tumefaciens (AgT-DHDPS). Enzyme-kinetics studies demonstrate that AgT-DHDPS possesses DHDPS activity in vitro. Crystals of AgT-DHDPS were grown in the unliganded form and in forms with substrate bound and with substrate plus allosteric inhibitor (lysine) bound. X-ray diffraction data sets were subsequently collected to a maximum resolution of 1.40 Å. Determination of the structure with and without substrate and inhibitor will offer insight into the design of novel pesticide agents.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Agrobacterium tumefaciens / enzymology*
  • Amino Acid Sequence
  • Cloning, Molecular
  • Crystallization
  • Gene Expression
  • Hydro-Lyases / chemistry*
  • Hydro-Lyases / genetics
  • Hydro-Lyases / isolation & purification
  • Isoenzymes / chemistry
  • Isoenzymes / genetics
  • Isoenzymes / isolation & purification
  • Ligands
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Structure, Quaternary
  • Sequence Alignment

Substances

  • Isoenzymes
  • Ligands
  • Hydro-Lyases
  • 4-hydroxy-tetrahydrodipicolinate synthase