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Manno-epi-cyclophellitols enable activity-based protein profiling of human α-mannosidases and discovery of new golgi mannosidase II inhibitors

DOI: 10.1021/jacs.0c03880 DOI Help

Authors: Zachary Armstrong (The University of York) , Chi-lin Kuo (Leiden University) , Daniël Lahav (Leiden University) , Bing Liu (Leiden University) , Rachel Johnson (The University of York) , Thomas J. M. Beenakker (Leiden University) , Casper De Boer (Leiden University) , Chung-sing Wong (Leiden University) , Erwin R. Van Rijssel (Leiden University) , Marjoke F. Debets (Leiden University) , Bogdan I. Florea (Leiden University) , Colin Hissink (Leiden University) , Rolf G. Boot (Leiden University) , Paul P. Geurink (Leiden University Medical Centre) , Huib Ovaa (Leiden University Medical Centre) , Mario Van Der Stelt (Leiden University) , Gijsbert M. Van Der Marel (Leiden University) , Jeroen D. C. Codée (Leiden University) , Johannes M. F. G. Aerts (Leiden University) , Liang Wu (The University of York) , Herman S. Overkleeft (Leiden University) , Gideon Davies (The University of York)
Co-authored by industrial partner: No

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

State: Published (Approved)
Published: July 2020
Diamond Proposal Number(s): 18598

Abstract: Golgi mannosidase II (GMII) catalyzes the sequential hydrolysis of two mannosyl residues from GlcNAcMan5GlcNAc2 to produce GlcNAcMan3GlcNAc2, the precursor for all complex N-glycans, including the branched N-glycans associated with cancer. Inhibitors of GMII are potential cancer therapeutics, but their usefulness is limited by off-target effects, which produce α-mannosidosis-like symptoms. Despite many structural and mechanistic studies of GMII, we still lack a potent and selective inhibitor of this enzyme. Here, we synthesized manno-epi-cyclophellitol epoxide and aziridines and demonstrate their covalent modification and time-dependent inhibition of GMII. Application of fluorescent manno-epi-cyclophellitol aziridine derivatives enabled activity-based protein profiling of α-mannosidases from both human cell lysate and mouse tissue extracts. Synthesized probes also facilitated a fluorescence polarization-based screen for dGMII inhibitors. We identified seven previously unknown inhibitors of GMII from a library of over 350 iminosugars and investigated their binding modalities through X-ray crystallography. Our results reveal previously unobserved inhibitor binding modes and promising scaffolds for the generation of selective GMII inhibitors.

Journal Keywords: Rodent models; Inhibitors; Inhibition; Peptides and proteins; Probes

Subject Areas: Chemistry, Biology and Bio-materials


Instruments: I03-Macromolecular Crystallography , I04-Macromolecular Crystallography