title: The purification, crystallization and preliminary X-ray diffraction analysis of dihydrodipicolinate synthase from Clostridium botulinum.
abstract: In recent years, dihydrodipicolinate synthase (DHDPS; EC 4.2.1.52) has received considerable attention from both mechanistic and structural viewpoints. This enzyme, which is part of the diaminopimelate pathway leading to lysine, couples (S)-aspartate-beta-semialdehyde with pyruvate via a Schiff base to a conserved active-site lysine. In this paper, the expression, purification, crystallization and preliminary X-ray diffraction analysis of DHDPS from Clostridium botulinum, an important bacterial pathogen, are presented. The enzyme was crystallized in a number of forms, predominantly using PEG precipitants, with the best crystal diffracting to beyond 1.9 A resolution and displaying P4(2)2(1)2 symmetry. The unit-cell parameters were a = b = 92.9, c = 60.4 A. The crystal volume per protein weight (V(M)) was 2.07 A(3) Da(-1), with an estimated solvent content of 41%. The structure of the enzyme will help guide the design of novel therapeutics against the C. botulinum pathogen.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Melbourne, Parkville, Victoria 3010, Australia. rdobson@unimelb.edu.au
authors.forename: Renwick C J
authors.initials: RC
authors.lastname: Dobson
authors.forename: Sarah C
authors.initials: SC
authors.lastname: Atkinson
authors.forename: Michael A
authors.initials: MA
authors.lastname: Gorman
authors.forename: Janet M
authors.initials: JM
authors.lastname: Newman
authors.forename: Michael W
authors.initials: MW
authors.lastname: Parker
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Perugini
mesh_terms: D003014:Clostridium botulinum / Q000201:enzymology* / Q000235:genetics; D003460:Crystallization; D006836:Hydro-Lyases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D014961:X-Ray Diffraction
chemical_list: D006836:Hydro-Lyases; C020040:4-hydroxy-tetrahydrodipicolinate synthase