Open AccessPurification, crystallization and preliminary X‐ray crystallographic studies of the Mycobacterium tuberculosis DNA gyrase CTD
Author(s) -
Darmon Amélie,
Piton Jérémie,
Roué Mélanie,
Petrella Stéphanie,
Aubry Alexandra,
Mayer Claudine
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111051888
Subject(s) - dna gyrase , dna , topoisomerase , ctd , protein subunit , crystallography , crystallization , mycobacterium tuberculosis , cleavage (geology) , stereochemistry , chemistry , biology , microbiology and biotechnology , biochemistry , tuberculosis , gene , escherichia coli , medicine , paleontology , oceanography , organic chemistry , pathology , fracture (geology) , geology
Mycobacterium tuberculosis DNA gyrase, a nanomachine involved in regulation of DNA topology, is the only type II topoisomerase present in this organism and hence is the sole target of fluoroquinolone in the treatment of tuberculosis. The C‐terminal domain (CTD) of the DNA gyrase A subunit possesses a unique feature, the ability to wrap DNA in a chiral manner, that plays an essential role during the catalytic cycle. A construct of 36 kDa corresponding to this domain has been overproduced, purified and crystallized. Diffraction data were collected to 1.55 Å resolution. Cleavage of the N‐terminal His tag was crucial for obtaining crystals. The crystals belonged to space group P 2 1 2 1 2 1 , with one molecule in the asymmetric unit and a low solvent content (33%). This is the first report of the crystallization and preliminary X‐ray diffraction studies of a DNA gyrase CTD from a species that contains one unique type II topoisomerase.