Open AccessMolecular cloning, overexpression, purification, crystallization and preliminary X‐ray diffraction studies of histidinol phosphate aminotransferase (HisC2) from Mycobacterium tuberculosis
Author(s) -
Nasir Nazia,
Vyas Rajan,
Chugh Chetna,
Ahangar Mohammad Syed,
Biswal Bichitra K.
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309111045386
Subject(s) - nitrilotriacetic acid , orthorhombic crystal system , polyethylene glycol , nuclear chemistry , chemistry , crystallization , crystallography , crystal structure , biochemistry , chelation , inorganic chemistry , organic chemistry
HisC2 from Mycobacterium tuberculosis was overexpressed in M. smegmatis and purified to homogeneity using nickel–nitrilotriacetic acid metal‐affinity and gel‐filtration chromatography. Diffraction‐quality crystals were grown using the hanging‐drop vapour‐diffusion technique from a condition consisting of 7 mg ml −1 HisC2 (in 20 m M Tris pH 8.8, 50 m M NaCl and 5% glycerol), 1 M succinic acid pH 7.0, 0.1 M HEPES pH 7.0 and 1%( w / v ) polyethylene glycol monomethyl ether 2000. The crystals belonged to the orthorhombic space group P 2 1 2 1 2, with unit‐cell parameters a = 255.98, b = 77.09, c = 117.97 Å. X‐ray diffraction data were recorded to 2.45 Å resolution from a single crystal using the in‐house X‐ray facility.