Cloning to crystallization of dihydrodipicolinate synthase from the intracellular pathogen Legionella pneumophila

• Published in: Acta crystallographica. Section F, Structural biology and crystallization communications Structural biology and crystallization communications, 2013-10, Vol.69 (10), p.1177-1181
• Main Authors: Siddiqui, Tanzeela, Paxman, Jason J., Dogovski, Con, Panjikar, Santosh, Perugini, Matthew A.
• Format: Article
• Language: English
• Subjects: Chains; Cloning; Cloning, Molecular; Crystal lattices; Crystallization; Crystallization Communications; Crystallography, X-Ray; dihydrodipicolinate synthase; Electrophoresis, Polyacrylamide Gel; Hydro-Lyases - chemistry; Hydro-Lyases - isolation & purification; Intracellular Space - parasitology; Legionella pneumophila; Legionella pneumophila - enzymology; Pathogens; Pseudomonas aeruginosa; Recombinant Proteins - chemistry; Recombinant Proteins - isolation & purification; Searching; Spectrometry, Mass, Electrospray Ionization
• ISSN: 1744-3091; 1744-3091; 2053-230X
• DOI: 10.1107/S1744309113024639

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 P6122, 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.