title: Expression, purification, crystallization and preliminary X-ray analysis of wild-type and of an active-site mutant of glyceraldehyde-3-phosphate dehydrogenase from Campylobacter jejuni.
abstract: The genome of the enteric pathogen Campylobacter jejuni encodes a single glyceraldehyde-3-phosphate dehydrogenase that can utilize either NADP+ or NAD+ as coenzymes for the oxidative phosphorylation of glyceraldehyde-3-phosphate to 1,3-diphosphoglycerate. Here, the cloning, expression, purification, crystallization and preliminary X-ray analysis of both the wild type and an active-site mutant of the enzyme are presented. Preliminary X-ray analysis revealed that in both cases the crystals diffracted to beyond 1.9 Å resolution. The space group is shown to be I4(1)22, with unit-cell parameters a=90.75, b=90.75, c=225.48 Å, α=90.46, β=90.46, γ=222.79°; each asymmetric unit contains only one subunit of the tetrameric enzyme.
authors.affiliation: Henry Wellcome Laboratories for Structural Biology, Henry Wellcome Building, University of Leicester, Leicester LE1 9HN, England.
authors.forename: David S
authors.initials: DS
authors.lastname: Tourigny
authors.forename: Paul R
authors.initials: PR
authors.lastname: Elliott
authors.forename: Louise J
authors.initials: LJ
authors.lastname: Edgell
authors.forename: Gregg M
authors.initials: GM
authors.lastname: Hudson
authors.forename: Peter C E
authors.initials: PC
authors.lastname: Moody
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D016123:Campylobacter jejuni / Q000201:enzymology*; D020134:Catalytic Domain; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000737:chemistry* / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008969:Molecular Sequence Data
chemical_list: D001426:Bacterial Proteins; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases