Journal article

Grappling with anisotropic data, pseudo-merohedral twinning and pseudo-translational noncrystallographic symmetry: A case study involving pyruvate kinase

KA Donovan, SC Atkinson, SA Kessans, F Peng, TF Cooper, MDW Griffin, GB Jameson, RCJ Dobson

Acta Crystallographica Section D Structural Biology | Published : 2016

Abstract

Pyruvate kinase is a key regulatory enzyme involved in the glycolytic pathway. The crystal structure of Escherichia coli type I pyruvate kinase was first solved in 1995 at 2.5 Å resolution. However, the space group was ambiguous, being either primitive orthorhombic (P212121)orC-centred orthorhombic (C2221). Here, the structure determination and refinement of E. coli type I pyruvate kinase to 2.28 Å resolution are presented. Using the same crystallization conditions as reported previously, the enzyme was found to crystallize in space group P21. Determination of the space group was complicated owing to anisotropic data, pseudo-translational noncrystallographic symmetry and the pseudo-merohed-r..

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University of Melbourne Researchers

Grants

Awarded by Army Research Office


Funding Acknowledgements

We thank Professor Andrea Mattevi (Universita di Pavia, Italy) for providing the pykF clone and also for valuable comments on the manuscript. We gratefully acknowledge the assistance of the MX beamline scientists at the Australian Synchrotron, Victoria, Australia. Parts of this research were undertaken at the MX1 beamline of the Australian Synchrotron. Travel to the Australian Synchrotron was supported by the New Zealand Synchrotron Group. RCJD acknowledges the following for funding support in part: (i) the Ministry of Business, Innovation and Employment (contract UOCX1208), (ii) the New Zealand Royal Society Marsden Fund (contract UOC1013) and (iii) the US Army Research Laboratory and US Army Research Office under contract/grant No. W911NF-11-1-0481. We especially thank Jackie Healy for her indefatigable technical support. Lastly, we thank the reviewers for their very helpful comments and suggestions.