Open AccessA novel noncovalent complex of chorismate mutase and DAHP synthase from Mycobacterium tuberculosis : protein purification, crystallization and X‐ray diffraction analysis
Author(s) -
Ökvist Mats,
Sasso Severin,
Roderer Kathrin,
Kast Peter,
Krengel Ute
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109035878
Subject(s) - chorismate mutase , crystallization , mycobacterium tuberculosis , mutase , crystallography , chemistry , enzyme , biochemistry , tuberculosis , biosynthesis , organic chemistry , medicine , pathology
Chorismate mutase catalyzes a key step in the shikimate‐biosynthetic pathway and hence is an essential enzyme in bacteria, plants and fungi. Mycobacterium tuberculosis contains two chorismate mutases, a secreted and an intracellular one, the latter of which (MtCM; Rv0948c; 90 amino‐acid residues; 10 kDa) is the subject of this work. Here are reported the gene expression, purification and crystallization of MtCM alone and of its complex with another shikimate‐pathway enzyme, DAHP synthase (MtDS; Rv2178c; 472 amino‐acid residues; 52 kDa), which has been shown to enhance the catalytic efficiency of MtCM. The MtCM–MtDS complex represents the first noncovalent enzyme complex from the common shikimate pathway to be structurally characterized. Soaking experiments with a transition‐state analogue are also reported. The crystals of MtCM and the MtCM–MtDS complex diffracted to 1.6 and 2.1 Å resolution, respectively.