Manno- epi -cyclophellitols Enable Activity-Based Protein Profiling of Human α-Mannosidases and Discovery of New Golgi Mannosidase II Inhibitors

Journal of the American Chemical Society, vol. 142, pp. 13021–13029

Abstract

Golgi mannosidase II (GMII) catalyzes the sequential hydrolysis of two mannosyl residues from GlcNAcMan 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 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.

Authors 20

1. Zachary Armstrong

   University of York

   Affiliation as printed

   Structural Biology Laboratory, Department of Chemistry

   The University of York

   Structural Biology Laboratory, Department of Chemistry, The University of York, York YO10 5DD, United Kingdom

2. C.J. Kuo Aachen

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

3. D. Lahav Aachen

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

4. Rachel D. Johnson

   University of York

   Affiliation as printed

   Structural Biology Laboratory, Department of Chemistry

   The University of York

   Structural Biology Laboratory, Department of Chemistry, The University of York, York YO10 5DD, United Kingdom

5. Thomas J. M. Beenakker Aachen

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

6. Casper de Boer Aachen

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

7. Erwin R. van Rijssel Aachen

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

8. Marjoke F. Debets Aachen

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

9. Bogdan I. Florea Aachen

   Leiden University

   Affiliation as printed

   Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

10. Colin Hissink Aachen

    Leiden University

    Affiliation as printed

    Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

11. Rolf G. Boot Aachen

    Leiden University

    Affiliation as printed

    Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

12. Paul P. Geurink Aachen Oncode Institute Department of Cell and Chemical Biology

    Leiden University · Leiden University Medical Center · Oncode Institute

    Affiliation as printed

    Oncode Institute & Department of Cell and Chemical Biology, Leiden University Medical Centre, Einthovenweg 20, 2333 ZC Leiden, The Netherlands

13. Huib Ovaa Aachen Oncode Institute Department of Cell and Chemical Biology

    Leiden University · Leiden University Medical Center · Oncode Institute

    Affiliation as printed

    Oncode Institute & Department of Cell and Chemical Biology, Leiden University Medical Centre, Einthovenweg 20, 2333 ZC Leiden, The Netherlands

14. Mario van der Stelt Aachen

    Leiden University

    Affiliation as printed

    Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

15. Gijsbert M. van der Marel Aachen

    Leiden University

    Affiliation as printed

    Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

16. Jeroen D. C. Codée Aachen

    Leiden University

    Affiliation as printed

    Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

17. Johannes M. F. G. Aerts Aachen

    Leiden University

    Affiliation as printed

    Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

18. Liang Wu

    University of York

    Affiliation as printed

    Structural Biology Laboratory, Department of Chemistry

    The University of York

    Structural Biology Laboratory, Department of Chemistry, The University of York, York YO10 5DD, United Kingdom

19. Herman S. Overkleeft corresponding Aachen

    Leiden University

    Affiliation as printed

    Leiden Institute of Chemistry, Leiden University, Einsteinweg 55, 2333 CC Leiden, The Netherlands

20. G.J. Davies corresponding

    University of York

    Affiliation as printed

    Structural Biology Laboratory, Department of Chemistry

    The University of York

    Structural Biology Laboratory, Department of Chemistry, The University of York, York YO10 5DD, United Kingdom