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Updated: May 7, 2026
Mycobacterium tuberculosis Extracellular Vesicle Enrichment through Size Exclusion Chromatography
Published on: May 19, 2022
Cloning, overexpression, purification, crystallization and preliminary X-ray diffraction analysis of Rv0241c (HtdX)
Rupam Biswas1, Debajyoti Dutta, Amit Kumar Das
1Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721 302, India.
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 P4₃2₁2 or P4₁2₁2, with unit-cell parameters a=b=63.67, c=140.88 Å. Both crystals contained one molecule in the asymmetric unit.
