Open AccessCloning to crystallization of dihydrodipicolinate synthase from the intracellular pathogen Legionella pneumophila
Author(s) -
Siddiqui Tanzeela,
Paxman Jason J.,
Dogovski Con,
Panjikar Santosh,
Perugini Matthew A.
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113024639
Subject(s) - legionella pneumophila , cloning (programming) , intracellular , pathogen , crystallization , microbiology and biotechnology , biology , legionella , atp synthase , chemistry , bacteria , biochemistry , genetics , gene , computer science , organic chemistry , programming language
Dihydrodipicolinate synthase (DHDPS) catalyses the rate‐limiting step in the biosynthesis of meso ‐diaminopimelate and lysine. Here, the cloning, expression, purification and crystallization of DHDPS from the intracellular pathogen Legionella pneumophila are described. Crystals grown in the presence of high‐molecular‐weight PEG precipitant and magnesium chloride were found to diffract beyond 1.65 Å resolution. The crystal lattice belonged to the hexagonal space group P 6 1 22, with unit‐cell parameters a = b = 89.31, c = 290.18 Å, and contained two molecules in the asymmetric unit. The crystal structure was determined by molecular replacement using a single chain of Pseudomonas aeruginosa DHDPS as the search model.