title: Cloning, expression, purification, crystallization and preliminary X-ray studies of a pyridoxine 5'-phosphate oxidase from Mycobacterium smegmatis.
abstract: Pyridoxine 5'-phosphate oxidases (PNPOxs) are known to catalyse the terminal step in pyridoxal 5'-phosphate biosynthesis in a flavin mononucleotide-dependent manner in humans and Escherichia coli. Recent reports of a putative PNPOx from Mycobacterium tuberculosis, Rv1155, suggest that the cofactor or catalytic mechanism may differ in Mycobacterium species. To investigate this, a putative PNPOx from M. smegmatis, Msmeg_3380, has been cloned. This enzyme has been recombinantly expressed in E. coli and purified to homogeneity. Good-quality crystals of selenomethionine-substituted Msmeg_3380 were obtained by the hanging-drop vapour-diffusion technique and diffracted to 1.2 A using synchrotron radiation.
authors.affiliation: CSIRO Entomology, Black Mountain, ACT 2601, Australia. colin.jackson@csiro.au
authors.forename: Colin J
authors.initials: CJ
authors.lastname: Jackson
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Taylor
authors.forename: David B
authors.initials: DB
authors.lastname: Tattersall
authors.forename: Nigel G
authors.initials: NG
authors.lastname: French
authors.forename: Paul D
authors.initials: PD
authors.lastname: Carr
authors.forename: David L
authors.initials: DL
authors.lastname: Ollis
authors.forename: Robyn J
authors.initials: RJ
authors.lastname: Russell
authors.forename: John G
authors.initials: JG
authors.lastname: Oakeshott
mesh_terms: D003001:Cloning, Molecular; D003460:Crystallization / Q000379:methods*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D020102:Mycobacterium smegmatis / Q000201:enzymology*; D011734:Pyridoxaminephosphate Oxidase / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D012645:Selenomethionine / Q000378:metabolism; D014961:X-Ray Diffraction / Q000379:methods
chemical_list: D012645:Selenomethionine; D011734:Pyridoxaminephosphate Oxidase