Open AccessCloning, overexpression, purification, crystallization and preliminary X‐ray diffraction analysis of Rv0241c (HtdX) from Mycobacterium tuberculosis H37Rv
Author(s) -
Biswas Rupam,
Dutta Debajyoti,
Das Amit Kumar
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113023452
Subject(s) - crystallization , operon , orthorhombic crystal system , mycobacterium tuberculosis , crystallography , chemistry , sodium , crystal (programming language) , crystal structure , gene , escherichia coli , biochemistry , tuberculosis , organic chemistry , computer science , programming language , medicine , pathology
Rv0241c (HtdX) is a putative (3 R )‐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 I 4 1 , 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 P 4 3 2 1 2 or P 4 1 2 1 2, with unit‐cell parameters a = b = 63.67, c = 140.88 Å. Both crystals contained one molecule in the asymmetric unit.