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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.

SUBMITTER: Atkinson SC

PROVIDER: S-EPMC3433193 | biostudies-literature | 2012 Sep

REPOSITORIES: biostudies-literature

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Cloning, expression, purification and crystallization of dihydrodipicolinate synthase from Agrobacterium tumefaciens.

Atkinson Sarah C SC Dogovski Con C Dobson Renwick C J RC Perugini Matthew A MA

Acta crystallographica. Section F, Structural biology and crystallization communications 20120830 Pt 9

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 substra ...[more]

PMID: 22949190

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The purification, crystallization and preliminary X-ray diffraction analysis of dihydrodipicolinate synthase from Clostridium botulinum.

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