title: Cloning, expression, purification and crystallization of dihydrodipicolinate synthase from Agrobacterium tumefaciens.
abstract: Dihydrodipicolinate synthase (DHDPS) catalyzes the first committed step of the lysine-biosynthesis pathway in bacteria, plants and some fungi. This study describes the cloning, expression, purification and crystallization of DHDPS (NP_354047.1) from the plant pathogen Agrobacterium tumefaciens (AgT-DHDPS). Enzyme-kinetics studies demonstrate that AgT-DHDPS possesses DHDPS activity in vitro. Crystals of AgT-DHDPS were grown in the unliganded form and in forms with substrate bound and with substrate plus allosteric inhibitor (lysine) bound. X-ray diffraction data sets were subsequently collected to a maximum resolution of 1.40 Å. Determination of the structure with and without substrate and inhibitor will offer insight into the design of novel pesticide agents.
authors.affiliation: Department of Biochemistry, La Trobe Institute for Molecular Science, La Trobe University, Melbourne, Victoria 3086, Australia.
authors.forename: Sarah C
authors.initials: SC
authors.lastname: Atkinson
authors.forename: Con
authors.initials: C
authors.lastname: Dogovski
authors.forename: Renwick C J
authors.initials: RC
authors.lastname: Dobson
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Perugini
mesh_terms: D016960:Agrobacterium tumefaciens / Q000201:enzymology*; D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D003460:Crystallization; D015870:Gene Expression; D006836:Hydro-Lyases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D007527:Isoenzymes / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D008024:Ligands; D008958:Models, Molecular; D008969:Molecular Sequence Data; D020836:Protein Structure, Quaternary; D016415:Sequence Alignment
chemical_list: D007527:Isoenzymes; D008024:Ligands; D006836:Hydro-Lyases; C020040:4-hydroxy-tetrahydrodipicolinate synthase