Manno-epi-cyclophellitols enable activity-based protein profiling of human α-mannosidases and discovery of new Golgi mannosidase II inhibitors
| Autor: | Zachary Armstrong, Gideon J. Davies, Chi-Lin Kuo, Erwin R. van Rijssel, Thomas J. M. Beenakker, Bogdan I. Florea, Mario van der Stelt, Gijsbert M. van der Marel, Rachel Johnson, D. Lahav, Bing Liu, Colin Hissink, Marjoke F. Debets, Jeroen D. C. Codée, C.S. Wong, Johannes M. F. G. Aerts, Paul P. Geurink, Casper de Boer, Huib Ovaa, Rolf G. Boot, Liang Wu, Herman S. Overkleeft |
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| Jazyk: | angličtina |
| Rok vydání: | 2020 |
| Předmět: |
chemistry.chemical_classification
Lysis Activity-based proteomics Epoxide General Chemistry Aziridine Golgi apparatus 010402 general chemistry 01 natural sciences Biochemistry Catalysis 0104 chemical sciences 3. Good health carbohydrates (lipids) chemistry.chemical_compound symbols.namesake Colloid and Surface Chemistry Enzyme chemistry symbols Mannosidases Fluorescence anisotropy |
| Zdroj: | Journal of the American Chemical Society, 142(30), 13021-13029 Journal of the American Chemical Society, 142(30), 13021-13029. AMER CHEMICAL SOC |
| Popis: | Golgi mannosidase II (GMII) catalyzes the sequential hydrolysis of two mannosyl residues from GIcNAc-Man(5)GlcNAc(2) to produce GlcNAcMan(3) GlcNAc(2), 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 alpha-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 alpha-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. |
| Databáze: | OpenAIRE |
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