Structural studies of a surface-entropy reduction mutant of O-GlcNAcase.

MALES, Alexandra and DAVIES, Gideon J (2019). Structural studies of a surface-entropy reduction mutant of O-GlcNAcase. Acta crystallographica. Section D, Structural biology, 75 (Pt 1), 70-78.

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Abstract

The enzyme O-GlcNAcase catalyses the removal of the O-GlcNAc co/post-translational modification in multicellular eukaryotes. The enzyme has become of acute interest given the intimate role of O-GlcNAcylation in tau modification and stability; small-molecular inhibitors of human O-GlcNAcase are under clinical assessment for the treatment of tauopathies. Given the importance of structure-based and mechanism-based inhibitor design for O-GlcNAcase, it was sought to test whether different crystal forms of the human enzyme could be achieved by surface mutagenesis. Guided by surface-entropy reduction, a Glu602Ala/Glu605Ala variant [on the Gly11-Gln396/Lys535-Tyr715 construct; Roth et al. (2017), Nature Chem. Biol. 13, 610-612] was obtained which led to a new crystal form of the human enzyme. An increase in crystal contacts stabilized disordered regions of the protein, enabling 88% of the structure to be modelled; only 83% was possible for the wild-type construct. Although the binding of the C-terminus was consistent with the wild type, Lys713 in monomer A was bound in the -1 subsite of the symmetry-related monomer A and the active sites of the B monomers were vacant. The new crystal form presents an opportunity for enhanced soaking experiments that are essential to understanding the binding mechanism and substrate specificity of O-GlcNAcase.

Item Type:	Article
Uncontrolled Keywords:	Humans; Binding Sites; Protein Binding; Substrate Specificity; Entropy; Mutant Proteins; beta-N-Acetylhexosaminidases; O-GlcNAc; O-GlcNAcase; crystallization; neurodegeneration; surface-entropy reduction; Binding Sites; Entropy; Humans; Mutant Proteins; Protein Binding; Substrate Specificity; beta-N-Acetylhexosaminidases
Identification Number:	https://doi.org/10.1107/s2059798318016595
Page Range:	70-78
SWORD Depositor:	Symplectic Elements
Depositing User:	Symplectic Elements
Date Deposited:	27 Sep 2023 14:20
Last Modified:	11 Oct 2023 11:30
URI:	https://shura.shu.ac.uk/id/eprint/32403

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