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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 Å resolution and belong to space group P21, with unit-cell parameters a = 44.36, b = 79.03, c = 52.85 Å, β = 109.11°. There are two molecules of molecular weight 21 948 Da per asymmetric unit. [ABSTRACT FROM AUTHOR]

The enzymes of the shikimate pathway are potential targets for the development of new therapies because they are essential for bacteria but absent from mammals. The last step in this pathway is performed by chorismate synthase (CS), which catalyzes the conversion of 5-enolpyruvylshikimate-3-phosphate to chorismate. Optimization of crystallization trials allowed the crystallization of homogeneous recombinant CS from Mycobacterium tuberculosis (MtCS). The crystals of MtCS belong to space group P6422 (or P6222) and diffract to 2.8 Å resolution, with unit-cell parameters a = b = 129.7, c = 156.8 Å. There are two molecules in the asymmetric unit. Molecular-replacement trials were not successful. Heavy-atom derivative screening is in progress. [ABSTRACT FROM AUTHOR]