Cloning, expression, purification and crystallization of dihydrodipicolinate synthase from the psychrophile Shewanella benthica
Jacinta M Wubben, Con Dogovski, Renwick CJ Dobson, Rachel Codd, Juliet A Gerrard, Michael W Parker, Matthew A Perugini
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS | INT UNION CRYSTALLOGRAPHY | Published : 2010
Dihydrodipicolinate synthase (DHDPS) is an oligomeric enzyme that catalyzes the first committed step of the lysine-biosynthesis pathway in plants and bacteria, which yields essential building blocks for cell-wall and protein synthesis. DHDPS is therefore of interest to drug-discovery research as well as to studies that probe the importance of quaternary structure to protein function, stability and dynamics. Accordingly, DHDPS from the psychrophilic (cold-dwelling) organism Shewanella benthica (Sb-DHDPS) was cloned, expressed, purified and crystallized. The best crystals of Sb-DHDPS were grown in 200 mM ammonium sulfate, 100 mM bis-tris pH 5.0-6.0, 23-26%(w/v) PEG 3350, 0.02%(w/v) sodium azid..View full abstract
Awarded by Defense Threat Reduction Agency (DTRA)
We would like to acknowledge the friendly staff of the Bio21 Collaborative Crystallization Centre (Bio21-C<SUP>3</SUP>) and the beamline scientists from the Australian Synchrotron. We would also like to thank all other members of the Perugini laboratory for helpful discussions during the preparation of this manuscript. Finally, we acknowledge the Defense Threat Reduction Agency (DTRA; Project ID AB07CBT004), the Hermon Slade Foundation, the Sir John and Lady Higgins Scholarship fund for providing a post-graduate scholarship for JMW and the Australian Research Council for providing a Future Fellowship for MAP and a Federation Fellowship for MWP.