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Discovery of Selective Small-Molecule Activators of a Bacterial Glycoside Hydrolase

DOI: 10.1002/anie.201407081 DOI Help
PMID: 25291993 PMID Help

Authors: John Darby (University of York) , Jens Landstrom (University of York) , Christian Roth (University of York) , Yuan He (University of York) , Gideon J. Davies (University of York) , Roderick E. Hubbard (University of York)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Angewandte Chemie International Edition , VOL 53 (49) , PAGES 13419 - 13423

State: Published (Approved)
Published: December 2014
Diamond Proposal Number(s): 1221

Open Access Open Access

Abstract: Fragment-based approaches are used routinely to discover enzyme inhibitors as cellular tools and potential therapeutic agents. There have been few reports, however, of the discovery of small-molecule enzyme activators. Herein, we describe the discovery and characterization of small-molecule activators of a glycoside hydrolase (a bacterial O-GlcNAc hydrolase). A ligand-observed NMR screen of a library of commercially available fragments identified an enzyme activator which yielded an approximate 90?% increase in kcat/KM?values (kcat=catalytic rate constant; KM=Michaelis constant). This compound binds to the enzyme in close proximity to the catalytic center. Evolution of the initial hits led to improved compounds that behave as nonessential activators effecting both KM?and Vmax?values (Vmax=maximum rate of reaction). The compounds appear to stabilize an active “closed” form of the enzyme. Such activators could offer an orthogonal alternative to enzyme inhibitors for perturbation of enzyme activity in?vivo, and could also be used for glycoside hydrolase activation in many industrial processes.

Journal Keywords: Enzyme Catalysis; Glycoside Hydrolase; Glycosylation;Inhibitors;Protein Structures

Subject Areas: Biology and Bio-materials


Instruments: I24-Microfocus Macromolecular Crystallography