Cloning, expression, purification and preliminary crystallographic data for Rv3214 (EntD), a predicted cofactor-dependent phosphoglycerate mutase from Mycobacterium tuberculosis.
The Mycobacterium tuberculosis open reading frame Rv3214, annotated as a cofactor-dependent phosphoglycerate mutase, has been cloned and expressed as an N-terminally His-tagged protein. Tagged, untagged and selenomethionine-labelled forms of Rv3214 (EntD) have been purified using nickel-affinity chromatography and gel filtration. The selenomethionine-labelled crystals diffracted to 2.15 A resolution and belong to space group P2(1), with unit-cell parameters a = 44.36, b = 79.03, c = 52.85 A, beta = 109.11 degrees. There are two molecules of molecular weight 21,948 Da per asymmetric unit.
Watkins
Harriet A
HA
Centre for Molecular Biodiscovery, School of Biological Sciences, University of Auckland, Private Bag 92019, Auckland, New Zealand. h.watkins@auckland.ac.nz
Yu
MinMin
M
Baker
Edward N
EN
D003001:Cloning, Molecular; D003067:Coenzymes / Q000378:metabolism*; D018360:Crystallography, X-Ray; D015870:Gene Expression; D008970:Molecular Weight; D009169:Mycobacterium tuberculosis / Q000201:enzymology*; D010736:Phosphoglycerate Mutase / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D012542:Scattering, Radiation
D003067:Coenzymes; D011993:Recombinant Fusion Proteins; D010736:Phosphoglycerate Mutase