Crystallization and preliminary X-ray analysis of dihydrodipicolinate synthase from Clostridium botulinum in the presence of its substrate pyruvate

SC Atkinson; RCJ Dobson; JM Newman; MA Gorman; C Dogovski; MW Parker; MA Perugini

Journal article

Crystallization and preliminary X-ray analysis of dihydrodipicolinate synthase from Clostridium botulinum in the presence of its substrate pyruvate

SC Atkinson, RCJ Dobson, JM Newman, MA Gorman, C Dogovski, MW Parker, MA Perugini

Acta Crystallographica Section F Structural Biology and Crystallization Communications | INT UNION CRYSTALLOGRAPHY | Published : 2009

DOI: 10.1107/S1744309108039018

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 Å 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 Å, β = 126.3°. The crystal volume per protein weight (VM) was 2.3 Å3 Da-1 (based on the presence of two monomers in the asymmetric unit), with an estimated solvent content of 46%. The high-resolution..

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University of Melbourne Researchers

Renwick Dobson Author

Michael Gorman Author

Michael Parker Author

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Grants

Awarded by Defense Threat Reduction Agency

Awarded by Australian Research Council

Funding Acknowledgements

We would like to thank Geoff Hogg from the Microbiological Diagnostic Unit (University of Melbourne, Australia) for providing genomic DNA from C. botulinum (Hall A strain). We would also like to acknowledge the Defense Threat Reduction Agency (Contract No. W911NF-07-1-0105) and the Australian Research Council (DP0770888 and LX0776388) for providing funding, an Australian postdoctoral fellowship for MAP and a Federation fellowship for MWP. We thank Tom Caradoc-Davies and Trevor Huyton for valued assistance at the Australian Synchrotron.