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Bicyclic picomolar OGA inhibitors enable chemoproteomic mapping of its endogenous post-translational modifications

DOI: 10.1021/jacs.1c10504 DOI Help

Authors: Manuel González-Cuesta (Universidad de Sevilla) , Peter Sidhu (Simon Fraser University; University of British Columbia) , Roger A. Ashmus (Simon Fraser University) , Alexandra Males (University of York) , Cameron Proceviat (Simon Fraser University) , Zarina Madden (Simon Fraser University) , Jason C. Rogalski (University of British Columbia) , Jil A. Busmann (Simon Fraser University) , Leonard J. Foster (University of British Columbia) , José M. García Fernández (nstituto de Investigaciones Químicas (IIQ), CSIC, Universidad de Sevilla) , Gideon J. Davies (The University of York) , Carmen Ortiz Mellet (Universidad de Sevilla) , David J. Vocadlo (Simon Fraser University)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Journal Of The American Chemical Society

State: Published (Approved)
Published: January 2022
Diamond Proposal Number(s): 18598

Abstract: Owing to its roles in human health and disease, the modification of nuclear, cytoplasmic, and mitochondrial proteins with O-linked N-acetylglucosamine residues (O-GlcNAc) has emerged as a topic of great interest. Despite the presence of O-GlcNAc on hundreds of proteins within cells, only two enzymes regulate this modification. One of these enzymes is O-GlcNAcase (OGA), a dimeric glycoside hydrolase that has a deep active site cleft in which diverse substrates are accommodated. Chemical tools to control OGA are emerging as essential resources for helping to decode the biochemical and cellular functions of the O-GlcNAc pathway. Here we describe rationally designed bicyclic thiazolidine inhibitors that exhibit superb selectivity and picomolar inhibition of human OGA. Structures of these inhibitors in complex with human OGA reveal the basis for their exceptional potency and show that they extend out of the enzyme active site cleft. Leveraging this structure, we create a high affinity chemoproteomic probe that enables simple one-step purification of endogenous OGA from brain and targeted proteomic mapping of its post-translational modifications. These data uncover a range of new modifications, including some that are less-known, such as O-ubiquitination and N-formylation. We expect that these inhibitors and chemoproteomics probes will prove useful as fundamental tools to decipher the mechanisms by which OGA is regulated and directed to its diverse cellular substrates. Moreover, the inhibitors and structures described here lay out a blueprint that will enable the creation of chemical probes and tools to interrogate OGA and other carbohydrate active enzymes.

Journal Keywords: Modification; Peptides and proteins; Inhibitors; Selectivity; Post-translational modification

Diamond Keywords: Enzymes

Subject Areas: Biology and Bio-materials, Chemistry


Instruments: I03-Macromolecular Crystallography , I04-1-Macromolecular Crystallography (fixed wavelength)

Added On: 10/01/2022 08:19

Discipline Tags:

Biochemistry Chemistry Structural biology Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)