title: Expression, purification and preliminary crystallographic analysis of dipeptidyl peptidase IV from Porphyromonas gingivalis.
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.
authors.affiliation: Department of Biological Sciences, University of Warwick, Gibbet Hill Road, Coventry CV4 7AL, England.
authors.forename: Dean
authors.initials: D
authors.lastname: Rea
authors.forename: Anne Marie
authors.initials: AM
authors.lastname: Lambeir
authors.forename: Yumi
authors.initials: Y
authors.lastname: Kumagai
authors.forename: Ingrid
authors.initials: I
authors.lastname: De Meester
authors.forename: Simon
authors.initials: S
authors.lastname: Scharpé
authors.forename: Vilmos
authors.initials: V
authors.lastname: Fülöp
mesh_terms: D002845:Chromatography; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D018819:Dipeptidyl Peptidase 4 / Q000737:chemistry*; D004926:Escherichia coli / Q000378:metabolism; D010957:Plasmids / Q000378:metabolism; D016966:Porphyromonas gingivalis / Q000201:enzymology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012645:Selenomethionine / Q000737:chemistry; D017356:Synchrotrons; D014961:X-Ray Diffraction
chemical_list: D011993:Recombinant Fusion Proteins; D012645:Selenomethionine; D018819:Dipeptidyl Peptidase 4