Cloning, expression, purification, crystallization and preliminary X-ray studies of a pyridoxine 5'-phosphate oxidase from Mycobacterium smegmatis.
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.
Jackson
Colin J
CJ
CSIRO Entomology, Black Mountain, ACT 2601, Australia. colin.jackson@csiro.au
Taylor
Matthew C
MC
Tattersall
David B
DB
French
Nigel G
NG
Carr
Paul D
PD
Ollis
David L
DL
Russell
Robyn J
RJ
Oakeshott
John G
JG
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
D012645:Selenomethionine; D011734:Pyridoxaminephosphate Oxidase