Journal article

Substrate Locking Promotes Dimer-Dimer Docking of an Enzyme Antibiotic Target

Sarah C Atkinson, Con Dogovski, Kathleen Wood, Michael DW Griffin, Michael A Gorman, Lilian Hor, Cyril F Reboul, Ashley M Buckle, Joachim Wuttke, Michael W Parker, Renwick Cj Dobson, Matthew A Perugini

Structure | CELL PRESS | Published : 2018


Protein dynamics manifested through structural flexibility play a central role in the function of biological molecules. Here we explore the substrate-mediated change in protein flexibility of an antibiotic target enzyme, Clostridium botulinum dihydrodipicolinate synthase. We demonstrate that the substrate, pyruvate, stabilizes the more active dimer-of-dimers or tetrameric form. Surprisingly, there is little difference between the crystal structures of apo and substrate-bound enzyme, suggesting protein dynamics may be important. Neutron and small-angle X-ray scattering experiments were used to probe substrate-induced dynamics on the sub-second timescale, but no significant changes were observ..

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Awarded by Australian Research Council

Awarded by National Health and Medical Research Council

Awarded by New Zealand Royal Society Marsden Fund

Awarded by US Army Research Office

Funding Acknowledgements

We acknowledge the support and assistance of the friendly staff at the CSIRO Collaborative Crystallisation Centre; Australian Synchrotron; and La Trobe University-Comprehensive Proteomics Platform. We acknowledge the Australian Research Council for project funding (DP150103313) and Future Fellowships to M.A.P. (FT0991245) and M.D.W.G. (FT140100544); infrastructure support from the Australian Cancer Research Foundation and the Victorian Government Operational Infrastructure Support Scheme to St. Vincent's Institute; the National Health and Medical Research Council for fellowship support for S.C.A. (1072267) and M.W.P. (1021645). Travel to perform experiments at FRM-II was funded by the access to major facilities program, supported by the Commonwealth of Australia under the International Science Linkages program. R.C.J.D. acknowledges the following for funding support, in part: (1) the New Zealand Royal Society Marsden Fund (contract UOC1506); (2) the US Army Research Laboratory and US Army Research Office under contract/grant number W911NF-11-1-0481; and (3) the Biomolecular Interactions Centre (UC).