Crystallization and preliminary X-ray diffraction analysis of diaminopimelate epimerase from Escherichia coli
Lilian Hor, Renwick CJ Dobson, Con Dogovski, Craig A Hutton, Matthew A Perugini
Acta Crystallographica Section F - Structural Biology and Crystallization Communications | INT UNION CRYSTALLOGRAPHY | Published : 2010
Diaminopimelate (DAP) epimerase (EC 18.104.22.168) catalyzes the penultimate step of lysine biosynthesis in bacteria and plants, converting L,L-diaminopimelate to meso-diaminopimelate. Here, the cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of DAP epimerase from Escherichia coli are presented. Crystals were obtained in space group P4(1)2(1)2 and diffracted to 2.0 A resolution, with unit-cell parameters a = b = 89.4, c = 179.6 A. Molecular replacement was conducted using Bacillus anthracis DAP epimerase as a search model and showed the presence of two molecules in the asymmetric unit, with an initial R(free) of 0.456 and R(work) of 0.416.
Awarded by Defense Threat Reduction Agency (DTRA)
We would firstly like to acknowledge the support and assistance of the friendly staff, especially Dr Janet Newman, at the Bio21 Collaborative Crystallographic Centre at CSIRO Molecular and Health Technologies, Parkville, Melbourne as well as all members of the Perugini and Hutton laboratories for helpful discussions during the preparation of this manuscript. This research was undertaken on the MX2 beamline 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. Finally, we acknowledge the Defense Threat Reduction Agency (DTRA; Project ID AB07CBT004) for program funding, the Australian Research Council for providing a Future Fellowship to MAP and The University of Melbourne for the C. R. Roper Research Fellowhip to RCJD.