Journal article

N-acetylmannosamine-6-phosphate 2-epimerase uses a novel substrate-assisted mechanism to catalyze amino sugar epimerization

Michael J Currie, Lavanyaa Manjunath, Christopher R Horne, Phillip M Rendle, Ramaswamy Subramanian, Rosmarie Friemann, Antony J Fairbanks, Andrew C Muscroft-Taylor, Rachel A North, Renwick CJ Dobson

JOURNAL OF BIOLOGICAL CHEMISTRY | ELSEVIER | Published : 2021

Abstract

There are five known general catalytic mechanisms used by enzymes to catalyze carbohydrate epimerization. The amino sugar epimerase N-acetylmannosamine-6-phosphate 2-epimerase (NanE) has been proposed to use a deprotonation-reprotonation mechanism, with an essential catalytic lysine required for both steps. However, the structural determinants of this mechanism are not clearly established. We characterized NanE from Staphylococcus aureus using a new coupled assay to monitor NanE catalysis in real time and found that it has kinetic constants comparable with other species. The crystal structure of NanE from Staphylococcus aureus, which comprises a triosephosphate isomerase barrel fold with an ..

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Grants

Awarded by New Zealand Royal Society Marsden Fund


Awarded by Ministry of Business, Innovation and Employment Smart Ideas grant


Funding Acknowledgements

We acknowledge the New Zealand Royal Society Marsden Fund (contracts UOC1506 and UOC1910 to R. C. J. D.) and the Ministry of Business, Innovation and Employment Smart Ideas grant (contract UOCX1706 to R. C. J. D.) . This research was supported by an AINSE Ltd Honours Scholarship to M. J. C.