Journal article

A tetrameric structure is not essential for activity in dihydrodipicolinate synthase (DHDPS) from Mycobacterium tuberculosis

Genevieve Evans, Linda Schuldt, Michael DW Griffin, Sean RA Devenish, F Grant Pearce, Matthew A Perugini, Renwick CJ Dobson, Geoffrey B Jameson, Manfred S Weiss, Juliet A Gerrard

ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS | ELSEVIER SCIENCE INC | Published : 2011

Abstract

Dihydrodipicolinate synthase (DHDPS) is a validated antibiotic target for which a new approach to inhibitor design has been proposed: disrupting native tetramer formation by targeting the dimer-dimer interface. In this study, rational design afforded a variant of Mycobacterium tuberculosis, Mtb-DHDPS-A204R, with disrupted quaternary structure. X-ray crystallography (at a resolution of 2.1Å) revealed a dimeric protein with an identical fold and active-site structure to the tetrameric wild-type enzyme. Analytical ultracentrifugation confirmed the dimeric structure in solution, yet the dimeric mutant has similar activity to the wild-type enzyme. Although the affinity for both substrates was som..

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Grants

Awarded by Foundation for Research Science and Technology


Awarded by Royal Society of New Zealand


Awarded by Australian Research Council


Funding Acknowledgements

We thank Dr. Marie Squire for assistance with mass spectrometry; Assoc. Prof. Emily Parker for useful discussions: Jackie Healy for exquisite technical assistance; and Dr. Andrew Muscroft-Taylor for synthesis of the aspartate semi-aldehyde. SRAD is grateful to the Foundation for Research Science and Technology for postdoctoral funding (Contract UOCX0603). R.C.J.D. acknowledges the C.R. Roper and The Royal Society of New Zealand Marsden Fund (Contract UOC1013) for support and M.A.P. for Future Fellowship support from the Australian Research Council. M.D.W.G. is the recipient of an Australian Research Council Post Doctoral Fellowship (Project number: DP110103528).