Cloning, overexpression, purification, crystallization and preliminary X‐ray diffraction analysis of Rv0241c (HtdX) from Mycobacterium tuberculosis H37Rv. Issue 10 (8th October 2013)

• Record Type: Journal Article
• Authors: Biswas, Rupam; Dutta, Debajyoti; Das, Amit Kumar
• Is Part Of: Acta crystallographica. Volume 69:Issue 10(2013:Oct.)
• Page Start: 1110
• Page End: 1113
• Publication Date: 2013-10-08
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S1744309113023452
• Languages: English
• ISSNs: 1744-3091

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>Rv0241c (HtdX) is a putative (3<italic>R</italic>)‐hydroxyacyl‐CoA dehydratase of <italic>Mycobacterium tuberculosis</italic>. The <italic>htdX</italic> 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 <italic>M</italic> sodium chloride, 0.1 <italic>M</italic> Na HEPES pH 8.0 and (ii) 2.5 <italic>M</italic> sodium chloride, 0.1 <italic>M</italic> 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 <italic>I</italic>4₁, with unit‐cell parameters <italic>a</italic> = <italic>b</italic> = 61.51, <italic>c</italic> = 143.81 Å. Crystals from the second condition diffracted to 3.1 Å resolution and belonged to space group <italic>P</italic>4₃2₁2 or <italic>P</italic>4₁2₁2, with unit‐cell parameters <italic>a</italic> = <italic>b</italic> = 63.67, <italic>c</italic> = 140.88 Å. Both crystals contained one molecule in the asymmetric unit.</p> </abstract>