title: Crystallization and preliminary X-ray analysis of dihydrodipicolinate synthase from Clostridium botulinum in the presence of its substrate pyruvate.
abstract: In this paper, the crystallization and preliminary X-ray diffraction analysis to near-atomic resolution of DHDPS from Clostridium botulinum crystallized in the presence of its substrate pyruvate are presented. The enzyme crystallized in a number of forms using a variety of PEG precipitants, with the best crystal diffracting to 1.2 A resolution and belonging to space group C2, in contrast to the unbound form, which had trigonal symmetry. The unit-cell parameters were a = 143.4, b = 54.8, c = 94.3 A, beta = 126.3 degrees . The crystal volume per protein weight (V(M)) was 2.3 A(3) Da(-1) (based on the presence of two monomers in the asymmetric unit), with an estimated solvent content of 46%. The high-resolution structure of the pyruvate-bound form of C. botulinum DHDPS will provide insight into the function and stability of this essential bacterial enzyme.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Melbourne, Parkville, Victoria, Australia.
authors.forename: Sarah C
authors.initials: SC
authors.lastname: Atkinson
authors.forename: Renwick C J
authors.initials: RC
authors.lastname: Dobson
authors.forename: Janet M
authors.initials: JM
authors.lastname: Newman
authors.forename: Michael A
authors.initials: MA
authors.lastname: Gorman
authors.forename: Con
authors.initials: C
authors.lastname: Dogovski
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*; D003460:Crystallization; D018360:Crystallography, X-Ray; D006836:Hydro-Lyases / Q000737:chemistry*; D019289:Pyruvic Acid / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D013379:Substrate Specificity
chemical_list: D011994:Recombinant Proteins; D019289:Pyruvic Acid; D006836:Hydro-Lyases; C020040:4-hydroxy-tetrahydrodipicolinate synthase