Open AccessExpression, purification and preliminary crystallographic analysis of dipeptidyl peptidase IV from Porphyromonas gingivalis
Author(s) -
Rea Dean,
Lambeir AnneMarie,
Kumagai Yumi,
De Meester Ingrid,
Scharpé Simon,
Fülöp Vilmos
Publication year - 2004
Publication title -
acta crystallographica section d
Language(s) - English
Resource type - Journals
ISSN - 1399-0047
DOI - 10.1107/s0907444904017639
Subject(s) - porphyromonas gingivalis , dipeptidyl peptidase , extracellular , chemistry , proteases , escherichia coli , crystallography , enzyme , nuclear chemistry , bacteria , biochemistry , biology , genetics , gene
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 m M Tris–HCl pH 8.0. Diffraction data to 2.7 Å resolution were collected using synchrotron radiation. The crystals belong to space group P 2 1 , with unit‐cell parameters a = 117.0, b = 112.9, c = 310.0 Å, β = 95.0°. 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.