Crystallization and preliminary X-ray diffraction analysis of L,L-diaminopimelate aminotransferase (DapL) from Chlamydomonas reinhardtii
Andre O Hudson, Irma Giron, Renwick CJ Dobson
ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY COMMUNICATIONS | INT UNION CRYSTALLOGRAPHY | Published : 2011
In the anabolic synthesis of diaminopimelate and lysine in plants and in some bacteria, the enzyme L,L-diaminopimelate aminotransferase (DapL; EC 184.108.40.206) catalyzes the conversion of tetrahydrodipicolinic acid (THDPA) to L,L-diaminopimelate, bypassing the DapD, DapC and DapE enzymatic steps in the bacterial acyl pathways. Here, the cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of DapL from the alga Chlamydomonas reinhardtii are presented. Protein crystals were grown in conditions containing 25% (w/v) PEG 3350 and 200 mM lithium sulfate and initially diffracted to ∼1.35 Å resolution. They belonged to space group P2(1)2(1)2(1), with unit-cell pa..View full abstract
Awarded by Office of Science of the US Department of Energy
We wish to thank the School of Biological and Medical Sciences at RIT for the support of this work through a Faculty Evaluation and Development (FEAD) 2010 award to AOH. IG is currently enrolled in the Biotechnology program at RIT. The DAP isomers used for enzymatic assays were a gift provided by Dr John Vederas of the University of Alberta, Canada. We would also 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. 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. RCJD acknowledges the C. R. Roper Bequest for Fellowship support. The work conducted by the US Department of Energy Joint Genome Institute is supported by the Office of Science of the US Department of Energy under Contract No. DE-AC02-05CH11231.