Amino Acids

, Volume 40, Issue 3, pp 829–839

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

Original Article

DOI: 10.1007/s00726-010-0700-6

Cite this article as:
He, Y., Bubb, A.K., Stubbs, K.A. et al. Amino Acids (2011) 40: 829. doi:10.1007/s00726-010-0700-6

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.

Keywords

Carbohydrate Enzyme X-ray structure O-GlcNAc Diabetes Neurodegeneration 

Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  1. 1.York Structural Biology Laboratory, Department of ChemistryThe University of YorkYorkUK
  2. 2.Department of ChemistrySimon Fraser UniversityBurnabyCanada
  3. 3.School of Biomedical, Biomolecular and Chemical SciencesThe University of Western Australia (M310)CrawleyAustralia

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