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Inhibition of a bacterial O-GlcNAcase homologue by lactone and lactam derivatives: structural, kinetic and thermodynamic analyses

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Abstract

The dynamic, intracellular, O-GlcNAc modification is of continuing interest and one whose study through targeted “chemical genetics” approaches is set to increase. Of particular importance is the inhibition of the O-GlcNAc hydrolase, O-GlcNAcase (OGA), since this provides a route to elevate cellular O-GlcNAc levels, and subsequent phenotypic evaluation. Such a small molecule approach complements other methods and potentially avoids changes in protein–protein interactions that manifest themselves in molecular biological approaches to O-GlcNAc transferase knockout or over-expression. Here we describe the kinetic, thermodynamic and three-dimensional structural analysis of a bacterial OGA analogue from Bacteroides thetaiotaomicron, BtGH84, in complex with a lactone oxime (LOGNAc) and a lactam form of N-acetylglucosamine and compare their binding signatures with that of the more potent inhibitor O-(2-acetamido-2-deoxy-d-glucopyranosylidene)amino N-phenyl carbamate (PUGNAc). We show that both LOGNAc and the N-acetyl gluconolactam are significantly poorer inhibitors than PUGNAc, which may reflect poorer mimicry of transition state geometry and steric clashes with the enzyme upon binding; drawbacks that the phenyl carbamate adornment of PUGNAc helps mitigate. Implications for the design of future generations of inhibitors are discussed.

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Acknowledgments

The authors thank the Biotechnology and Biological Sciences Research Council for their funding. G. J. D. is a Royal Society Wolfson Research Merit Award recipient. Y. H. was partially supported by the Wild Fund of the Department of Chemistry at York. T. M. G. is a Sir Henry Wellcome Postdoctoral Fellow, funded by the Wellcome Trust. Professor Andrea Vasella (ETH Zürich) is thanked for provision of N-acetyl gluconolactam 3.

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  1. Tracey M. Gloster

    Present address: Department of Chemistry, Simon Fraser University, 8888 University Drive, Burnaby, BC, V5A 1S6, Canada

  2. Keith A. Stubbs

    Present address: School of Biomedical, Biomolecular and Chemical Sciences, The University of Western Australia (M310), 35 Stirling Highway, Crawley, WA, 6009, Australia

Authors and Affiliations

  1. York Structural Biology Laboratory, Department of Chemistry, The University of York, York, YO10 5DD, UK

    Yuan He, Abigail K. Bubb, Tracey M. Gloster & Gideon J. Davies

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  1. Yuan He
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Correspondence to Tracey M. Gloster or Gideon J. Davies.

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He, Y., Bubb, A.K., Stubbs, K.A. et al. Inhibition of a bacterial O-GlcNAcase homologue by lactone and lactam derivatives: structural, kinetic and thermodynamic analyses. Amino Acids 40, 829–839 (2011). https://doi.org/10.1007/s00726-010-0700-6

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