A novel noncovalent complex of chorismate mutase and DAHP synthase from Mycobacterium tuberculosis: protein purification, crystallization and X-ray diffraction analysis.
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 A resolution, respectively.
Department of Chemistry, University of Oslo, NO-0315 Oslo, Norway.
Mats
M
Okvist
Severin
S
Sasso
Kathrin
K
Roderer
Peter
P
Kast
Ute
U
Krengel
D010723:3-Deoxy-7-Phosphoheptulonate Synthase / Q000737:chemistry*; D002826:Chorismate Mutase / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D009097:Multienzyme Complexes / Q000737:chemistry; D009169:Mycobacterium tuberculosis / Q000201:enzymology*
D009097:Multienzyme Complexes; D010723:3-Deoxy-7-Phosphoheptulonate Synthase; D002826:Chorismate Mutase