title: Overproduction and preliminary crystallographic study of a human kynurenine aminotransferase II homologue from Pyrococcus horikoshii OT3.
abstract: The Pyrococcus horikoshii OT3 genome contains a gene encoding a human kynurenine aminotransferase II (KAT II) homologue, which consists of 428 amino-acid residues and shows an amino-acid sequence identity of 30% to human KAT II. This gene was overexpressed in Escherichia coli and the recombinant protein (Ph-KAT II) was purified. Gel-filtration chromatography showed that Ph-KAT II exists as a homodimer. Ph-KAT II exhibited enzymatic activity that catalyzes the transamination of L-kynurenine to produce kynurenic acid. Crystals of Ph-KAT II were grown using the sitting-drop vapour-diffusion method and native X-ray diffraction data were collected to 2.2 A resolution using synchrotron radiation from station BL44XU at SPring-8. The crystals belong to the centred orthorhombic space group C222(1), with unit-cell parameters a = 71.75, b = 86.84, c = 137.30 A. Assuming one molecule per asymmetric unit, the VM value was 2.19 A3 Da(-1) and the solvent content was 43.3%.
authors.affiliation: Department of Material and Life Science, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan.
authors.forename: Hyongi
authors.initials: H
authors.lastname: Chon
authors.forename: Hiroyoshi
authors.initials: H
authors.lastname: Matsumura
authors.forename: Shoko
authors.initials: S
authors.lastname: Shimizu
authors.forename: Nao
authors.initials: N
authors.lastname: Maeda
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Koga
authors.forename: Kazufumi
authors.initials: K
authors.lastname: Takano
authors.forename: Shigenori
authors.initials: S
authors.lastname: Kanaya
mesh_terms: D019843:Archaeal Proteins / Q000737:chemistry / Q000302:isolation & purification; D001483:Base Sequence; D003460:Crystallization; D017931:DNA Primers; D020745:Genome, Archaeal; D006801:Humans; D043902:Pyrococcus horikoshii / Q000201:enzymology* / Q000235:genetics; D021241:Spectrometry, Mass, Electrospray Ionization; D000637:Transaminases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D014162:Transfection; D014961:X-Ray Diffraction
chemical_list: D019843:Archaeal Proteins; D017931:DNA Primers; D000637:Transaminases; C021481:kynurenine-oxoglutarate transaminase