Open AccessSample preparation, crystallization and structure solution of HisC from Mycobacterium tuberculosis
Author(s) -
Nasir Nazia,
Vyas Rajan,
Biswal Bichitra K.
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113006210
Subject(s) - monoclinic crystal system , polyethylene glycol , materials science , crystallization , mycobacterium tuberculosis , chromatography , crystallography , nitrilotriacetic acid , corynebacterium glutamicum , chemistry , crystal structure , chelation , inorganic chemistry , biochemistry , tuberculosis , organic chemistry , medicine , pathology , gene
Histidinolphosphate aminotransferase (HisC; Rv1600) 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 suitable for X‐ray analysis were grown by the hanging‐drop vapour‐diffusion technique using 30% polyethylene glycol monomethyl ether 2000 as the precipitant. The crystals belonged to the hexagonal space group P 3 2 21, with an unusual high solvent content of 74.5%. X‐ray diffraction data were recorded to 3.08 Å resolution from a single crystal using in‐house Cu K α radiation. The structure of HisC was solved by the molecular‐replacement method using its Corynebacterium glutamicum counterpart as a search model. HisC is a dimer in the crystal as well as in solution.