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

TY - JOUR

T1 - Manno- epi-cyclophellitols Enable Activity-Based Protein Profiling of Human α-Mannosidases and Discovery of New Golgi Mannosidase II Inhibitors

AU - Armstrong, Zachary

AU - Kuo, Chi Lin

AU - Lahav, Daniël

AU - Liu, Bing

AU - Johnson, Rachel

AU - Beenakker, Thomas J.M.

AU - De Boer, Casper

AU - Wong, Chung Sing

AU - Van Rijssel, Erwin R.

AU - Debets, Marjoke F.

AU - Florea, Bogdan I.

AU - Hissink, Colin

AU - Boot, Rolf G.

AU - Geurink, Paul P.

AU - Ovaa, Huib

AU - Van Der Stelt, Mario

AU - Van Der Marel, Gijsbert M.

AU - Codée, Jeroen D.C.

AU - Aerts, Johannes M.F.G.

AU - Wu, Liang

AU - Overkleeft, Herman S.

AU - Davies, Gideon J.

PY - 2020/7/29

Y1 - 2020/7/29

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85089608765&partnerID=8YFLogxK

U2 - 10.1021/jacs.0c03880

DO - 10.1021/jacs.0c03880

M3 - Article

C2 - 32605368

AN - SCOPUS:85089608765

SN - 0002-7863

VL - 142

SP - 13021

EP - 13029

JO - Journal of the American Chemical Society

JF - Journal of the American Chemical Society

IS - 30

ER -