Cloning, overexpression, purification, crystallization and preliminary X-ray diffraction analysis of Rv0241c (HtdX) from Mycobacterium tuberculosis H37Rv.

ABSTRACT: Rv0241c (HtdX) is a putative (3R)-hydroxyacyl-CoA dehydratase of Mycobacterium tuberculosis. The htdX gene belongs to a conserved operon and is expressed in mycobacteria in the presence of several fatty-acid synthase II drugs. To elucidate the structure of HtdX, the protein was cloned, overexpressed, purified to homogeneity and crystallized. The protein was crystallized from two conditions: (i) 3?M sodium chloride, 0.1?M Na HEPES pH 8.0 and (ii) 2.5?M sodium chloride, 0.1?M Tris-HCl pH 8.5. A complete diffraction data set was collected from crystals from both conditions. The crystal from the first condition diffracted to 2.3?Å resolution and belonged to space group I41, with unit-cell parameters a=b=61.51, c=143.81?Å. Crystals from the second condition diffracted to 3.1?Å resolution and belonged to space group P4?2?2 or P4?2?2, with unit-cell parameters a=b=63.67, c=140.88?Å. Both crystals contained one molecule in the asymmetric unit.

SUBMITTER: Biswas R 

PROVIDER: S-EPMC3792668 | biostudies-literature | 2013 Oct

REPOSITORIES: biostudies-literature