Dennis, Rebecca J. and Taylor, Edward J. and Macauley, Matthew S. and Stubbs, Keith A. and Turkenberg, Johan P. and Hart, Samuel J. and Black, Gary N. and Vocadlo, David J. and Davies, Gideon J. (2006) Structure and mechanism of a bacterial beta-glucosaminidase having O-GlcNAcase activity. Nature Structural & Molecular Biology, 13 (4). pp. 365-371. ISSN 1545-9993

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Abstract

O-GlcNAc is an abundant post-translational modification of serine and threonine residues of nucleocytoplasmic proteins. This modification, found only within higher eukaryotes, is a dynamic modification that is often reciprocal to phosphorylation. In a manner analogous to phosphatases, a glycoside hydrolase termed O-GlcNAcase cleaves O-GlcNAc from modified proteins. Enzymes with high sequence similarity to human O-GlcNAcase are also found in human pathogens and symbionts. We report the three-dimensional structure of O-GlcNAcase from the human gut symbiont Bacteroides thetaiotaomicron both in its native form and in complex with a mimic of the reaction intermediate. Mutagenesis and kinetics studies show that the bacterial enzyme, very similarly to its human counterpart, operates via an unusual 'substrate-assisted' catalytic mechanism, which will inform the rational design of enzyme inhibitors. Dennis, R. J., Taylor, E. J., Macauley, M. S., Stubbs, K. A., Turkenburg, J. P., Hart, S. J., Black, G. N., Vocadlo, D. J. &Vocadlo, G. J.

Item Type:	Article
Additional Information:	O-GlcNAc is an abundant post-translational modification of serine and threonine residues of nucleocytoplasmic proteins. This modification, found only within higher eukaryotes, is a dynamic modification that is often reciprocal to phosphorylation. In a manner analogous to phosphatases, a glycoside hydrolase termed O-GlcNAcase cleaves O-GlcNAc from modified proteins. Enzymes with high sequence similarity to human O-GlcNAcase are also found in human pathogens and symbionts. We report the three-dimensional structure of O-GlcNAcase from the human gut symbiont Bacteroides thetaiotaomicron both in its native form and in complex with a mimic of the reaction intermediate. Mutagenesis and kinetics studies show that the bacterial enzyme, very similarly to its human counterpart, operates via an unusual 'substrate-assisted' catalytic mechanism, which will inform the rational design of enzyme inhibitors. Dennis, R. J., Taylor, E. J., Macauley, M. S., Stubbs, K. A., Turkenburg, J. P., Hart, S. J., Black, G. N., Vocadlo, D. J. &Vocadlo, G. J.
Keywords:	O-GlcNAcase, gh84
Subjects:	C Biological Sciences > C700 Molecular Biology, Biophysics and Biochemistry
Divisions:	College of Sciences > Faculty of Science > School of Life Sciences
Depositing User:	Edward Taylor
Date Deposited:	18 Sep 2012 19:25
Last Modified:	13 Mar 2013 09:13
URI:	http://eprints.lincoln.ac.uk/id/eprint/6167

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