Journal article
Cloning, expression, purification and crystallization of dihydrodipicolinate synthase from the grapevine Vitis vinifera
SC Atkinson, C Dogovski, J Newman, RCJ Dobson, MA Perugini
Acta Crystallographica Section F Structural Biology and Crystallization Communications | Published : 2011
Abstract
Dihydrodipicolinate synthase (DHDPS) catalyses 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 from the grapevine Vitis vinifera (Vv-DHDPS). Following in-drop cleavage of the hexahistidine tag, cocrystals of Vv-DHDPS with the substrate pyruvate were grown in 0.1 M Bis-Tris propane pH 8.2, 0.2 M sodium bromide, 20%(w/v) PEG 3350. X-ray diffraction data in space group P1 at a resolution of 2.2 Å are presented. Preliminary diffraction data analysis indicated the presence of eight molecules per asymmetric unit (V M = 2.55 Å 3 Da -1, 52% solvent content). The pen..
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Awarded by University of Melbourne
Awarded by New Zealand Royal Society
Funding Acknowledgements
We would firstly like to acknowledge the support and assistance of the friendly staff at the CSIRO Collaborative Crystallization Centre at CSIRO Material Science and Engineering, Parkville, Melbourne, Australia and the MX beamline scientists at the Australian Synchrotron, Victoria, Australia. The views expressed herein are those of the authors and are not necessarily those of the owner or operator of the Australian Synchrotron. We would also like to thank all members of the Perugini laboratory for helpful discussions during the preparation of this manuscript. Finally, we acknowledge the Australian Research Council for providing a Future Fellowship for MAP and The University of Melbourne for providing project funding (FRGSS 2011 project grant). RCJD acknowledges the C. R. Roper Bequest for fellowship support and the New Zealand Royal Society Marsden Fund for funding support (UOC1013).