Expression, purification and preliminary crystallographic analysis of dipeptidyl peptidase IV from Porphyromonas gingivalis

Acta Crystallogr D Biol Crystallogr. 2004 Oct;60(Pt 10):1871-3. doi: 10.1107/S0907444904017639. Epub 2004 Sep 23.

Abstract

The asaccharolytic periodontopathogen Porphyromonas gingivalis produces membrane-anchored proteases such as dipeptidyl peptidase IV that are involved in the destruction of host periodontal tissue. The extracellular domain of this enzyme was overexpressed in Escherichia coli as an N-terminal His-tag fusion protein, purified using standard metal-affinity chromatography and crystallized using the hanging-drop vapour-diffusion technique in 40% 2-methyl-2,4-pentanediol and 100 mM Tris-HCl pH 8.0. Diffraction data to 2.7 A resolution were collected using synchrotron radiation. The crystals belong to space group P2(1), with unit-cell parameters a = 117.0, b = 112.9, c = 310.0 A, beta = 95.0 degrees. There are ten molecules per asymmetric unit, indicating a solvent content of 50%. Data were also collected from selenomethionine-derived crystals and structure solution by SAD or MAD is in progress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography
  • Cloning, Molecular
  • Crystallization
  • Crystallography, X-Ray
  • Dipeptidyl Peptidase 4 / chemistry*
  • Escherichia coli / metabolism
  • Plasmids / metabolism
  • Porphyromonas gingivalis / enzymology*
  • Recombinant Fusion Proteins / chemistry
  • Selenomethionine / chemistry
  • Synchrotrons
  • X-Ray Diffraction

Substances

  • Recombinant Fusion Proteins
  • Selenomethionine
  • Dipeptidyl Peptidase 4