Purification, crystallisation and preliminary structural studies of dTDP-6-deoxy-D-xylo-4-hexulose 3,5 epimerase (RmlC) from Salmonella entrica serovar Typhimurim, the third enzyme of the dTDP-rhanose synthesis pathway

MF Giraud, FM Gordon, C Whitfield, P Messner, SA McMahon, James Henderson Naismith

Research output: Contribution to journalArticlepeer-review

21 Citations (Scopus)

Abstract

L-Rhamnose is an essential component of the cell wall of many pathogenic bacteria. Its precusor, dTDP-L-rhamnose, is synthesized from alpha-D-glucose-1-phosphate and dTTP via a pathway requiring four distinct enzymes: RmlA, RmlB, RmlC and RmlD. RmlC was overexpressed in Escherichia coli. The recombinant protein was purified by a two-step protocol involving anion-exchange and hydrophobic chromatography. Dynamic light-scattering experiments indicated that the recombinant protein is monodisperse. Crystals were obtained using the sitting-drop vapour-diffusion method with ammonium sulfate as precipitant. Diffraction data were collected on a frozen crystal to a resolution of 2.17 Angstrom. The crystal belongs to either space group P3(1)21 or P3(2)21, with unit-cell parameters a = b = 71.56, c = 183.53 Angstrom and alpha = beta = 90, gamma = 120 degrees.

Original languageEnglish
Pages (from-to)706-708
Number of pages3
JournalActa Crystallographica. Section D, Biological crystallography
VolumeD55
Publication statusPublished - Mar 1999

Keywords

  • GENE-CLUSTER
  • O-ANTIGEN
  • BIOSYNTHESIS
  • SEQUENCE
  • LIPOPOLYSACCHARIDE
  • BINDING
  • REGION

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