Structure of human endo-a-1,2-mannosidase (MANEA), an antiviral host-glycosylation target

Lukasz F. Sobala; Pearl Z. Fernandes; Zalihe Hakki; Andrew J. Thompson; Jonathon D. Howe; Michelle Hill; Nicole Zitzmann; Scott Davies; Zania Stamataki; Terry D. Butters; Dominic S. Alonzi; Spencer J. Williams; Gideon J. Davies

doi:10.1073/pnas.2013620117

Structure of human endo-a-1,2-mannosidase (MANEA), an antiviral host-glycosylation target

Lukasz F. Sobala, Pearl Z. Fernandes, Zalihe Hakki, Andrew J. Thompson, Jonathon D. Howe, Michelle Hill, Nicole Zitzmann, Scott Davies, Zania Stamataki, Terry D. Butters, Dominic S. Alonzi, Spencer J. Williams, Gideon J. Davies^*

^*Corresponding author for this work

Chemistry

Research output: Contribution to journal › Article › peer-review

Abstract

Mammalian protein N-linked glycosylation is critical for glycoprotein folding, quality control, trafficking, recognition, and function. N-linked glycans are synthesized from Glc₃Man₉GlcNAc₂precursors that are trimmed and modified in the endoplasmic reticulum (ER) and Golgi apparatus by glycoside hydrolases and glycosyltransferases. Endo-a-1,2-mannosidase (MANEA) is the sole endoacting glycoside hydrolase involved in N-glycan trimming and is located within the Golgi, where it allows ER-escaped glycoproteins to bypass the classical N-glycosylation trimming pathway involving ER glucosidases I and II. There is considerable interest in the use of small molecules that disrupt N-linked glycosylation as therapeutic agents for diseases such as cancer and viral infection. Here we report the structure of the catalytic domain of human MANEA and complexes with substrate-derived inhibitors, which provide insight into dynamic loop movements that occur on substrate binding. We reveal structural features of the human enzyme that explain its substrate preference and the mechanistic basis for catalysis. These structures have inspired the development of new inhibitors that disrupt host protein N-glycan processing of viral glycans and reduce the infectivity of bovine viral diarrhea and dengue viruses in cellular models. These results may contribute to efforts aimed at developing broad-spectrum antiviral agents and help provide a more in-depth understanding of the biology of mammalian glycosylation.

Original language	English
Pages (from-to)	29595-29601
Number of pages	7
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	117
Issue number	47
Early online date	5 Nov 2020
DOIs	https://doi.org/10.1073/pnas.2013620117
Publication status	Published - 24 Nov 2020

Bibliographical note

Funding Information:
ACKNOWLEDGMENTS. We thank Jon Agirre for assistance with generating a dictionary file for TEW, Eleanor Dodson for assistance with processing the highly anisotropic datasets, Alexandra Males for collecting ITC data with Glc-DMJ, and Mahima Sharma for assisting with supplemental figure production. We also thank Diamond Light Source UK for access to beamlines I03, I04, and I24 (Proposals mx1221, mx12587, and mx18598). Support for this work was provided by the European Research Council (ERC-2012-AdG-32294, “Glycopoise”), the Australian Research Council (DP120101396, FT130100103, and DP180101957), and the Royal Society (a Ken Murray Research Professorship, to G.J.D.).

Publisher Copyright:
© 2020 National Academy of Sciences. All rights reserved.

Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.

Keywords

Antiviral
Enzyme
Glycosylation
Secretory pathway
Structural biology

Access to Document

10.1073/pnas.2013620117

29595.fullFinal published version, 1.3 MBLicence: CC BY-NC-ND

Cite this

Sobala, L. F., Fernandes, P. Z., Hakki, Z., Thompson, A. J., Howe, J. D., Hill, M., Zitzmann, N., Davies, S., Stamataki, Z., Butters, T. D., Alonzi, D. S., Williams, S. J., & Davies, G. J. (2020). Structure of human endo-a-1,2-mannosidase (MANEA), an antiviral host-glycosylation target. Proceedings of the National Academy of Sciences of the United States of America, 117(47), 29595-29601. https://doi.org/10.1073/pnas.2013620117

@article{de2fb5c2f5644df39afa85b9b20d8d36,

title = "Structure of human endo-a-1,2-mannosidase (MANEA), an antiviral host-glycosylation target",

abstract = "Mammalian protein N-linked glycosylation is critical for glycoprotein folding, quality control, trafficking, recognition, and function. N-linked glycans are synthesized from Glc3Man9GlcNAc2precursors that are trimmed and modified in the endoplasmic reticulum (ER) and Golgi apparatus by glycoside hydrolases and glycosyltransferases. Endo-a-1,2-mannosidase (MANEA) is the sole endoacting glycoside hydrolase involved in N-glycan trimming and is located within the Golgi, where it allows ER-escaped glycoproteins to bypass the classical N-glycosylation trimming pathway involving ER glucosidases I and II. There is considerable interest in the use of small molecules that disrupt N-linked glycosylation as therapeutic agents for diseases such as cancer and viral infection. Here we report the structure of the catalytic domain of human MANEA and complexes with substrate-derived inhibitors, which provide insight into dynamic loop movements that occur on substrate binding. We reveal structural features of the human enzyme that explain its substrate preference and the mechanistic basis for catalysis. These structures have inspired the development of new inhibitors that disrupt host protein N-glycan processing of viral glycans and reduce the infectivity of bovine viral diarrhea and dengue viruses in cellular models. These results may contribute to efforts aimed at developing broad-spectrum antiviral agents and help provide a more in-depth understanding of the biology of mammalian glycosylation.",

keywords = "Antiviral, Enzyme, Glycosylation, Secretory pathway, Structural biology",

author = "Sobala, {Lukasz F.} and Fernandes, {Pearl Z.} and Zalihe Hakki and Thompson, {Andrew J.} and Howe, {Jonathon D.} and Michelle Hill and Nicole Zitzmann and Scott Davies and Zania Stamataki and Butters, {Terry D.} and Alonzi, {Dominic S.} and Williams, {Spencer J.} and Davies, {Gideon J.}",

note = "Funding Information: ACKNOWLEDGMENTS. We thank Jon Agirre for assistance with generating a dictionary file for TEW, Eleanor Dodson for assistance with processing the highly anisotropic datasets, Alexandra Males for collecting ITC data with Glc-DMJ, and Mahima Sharma for assisting with supplemental figure production. We also thank Diamond Light Source UK for access to beamlines I03, I04, and I24 (Proposals mx1221, mx12587, and mx18598). Support for this work was provided by the European Research Council (ERC-2012-AdG-32294, “Glycopoise”), the Australian Research Council (DP120101396, FT130100103, and DP180101957), and the Royal Society (a Ken Murray Research Professorship, to G.J.D.). Publisher Copyright: {\textcopyright} 2020 National Academy of Sciences. All rights reserved. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.",

year = "2020",

month = nov,

day = "24",

doi = "10.1073/pnas.2013620117",

language = "English",

volume = "117",

pages = "29595--29601",

journal = "Proceedings of the National Academy of Sciences of the United States of America",

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Sobala, LF, Fernandes, PZ, Hakki, Z, Thompson, AJ, Howe, JD, Hill, M, Zitzmann, N, Davies, S, Stamataki, Z, Butters, TD, Alonzi, DS, Williams, SJ & Davies, GJ 2020, 'Structure of human endo-a-1,2-mannosidase (MANEA), an antiviral host-glycosylation target', Proceedings of the National Academy of Sciences of the United States of America, vol. 117, no. 47, pp. 29595-29601. https://doi.org/10.1073/pnas.2013620117

TY - JOUR

T1 - Structure of human endo-a-1,2-mannosidase (MANEA), an antiviral host-glycosylation target

AU - Sobala, Lukasz F.

AU - Fernandes, Pearl Z.

AU - Hakki, Zalihe

AU - Thompson, Andrew J.

AU - Howe, Jonathon D.

AU - Hill, Michelle

AU - Zitzmann, Nicole

AU - Davies, Scott

AU - Stamataki, Zania

AU - Butters, Terry D.

AU - Alonzi, Dominic S.

AU - Williams, Spencer J.

AU - Davies, Gideon J.

N1 - Funding Information: ACKNOWLEDGMENTS. We thank Jon Agirre for assistance with generating a dictionary file for TEW, Eleanor Dodson for assistance with processing the highly anisotropic datasets, Alexandra Males for collecting ITC data with Glc-DMJ, and Mahima Sharma for assisting with supplemental figure production. We also thank Diamond Light Source UK for access to beamlines I03, I04, and I24 (Proposals mx1221, mx12587, and mx18598). Support for this work was provided by the European Research Council (ERC-2012-AdG-32294, “Glycopoise”), the Australian Research Council (DP120101396, FT130100103, and DP180101957), and the Royal Society (a Ken Murray Research Professorship, to G.J.D.). Publisher Copyright: © 2020 National Academy of Sciences. All rights reserved. Copyright: Copyright 2020 Elsevier B.V., All rights reserved.

PY - 2020/11/24

Y1 - 2020/11/24

N2 - Mammalian protein N-linked glycosylation is critical for glycoprotein folding, quality control, trafficking, recognition, and function. N-linked glycans are synthesized from Glc3Man9GlcNAc2precursors that are trimmed and modified in the endoplasmic reticulum (ER) and Golgi apparatus by glycoside hydrolases and glycosyltransferases. Endo-a-1,2-mannosidase (MANEA) is the sole endoacting glycoside hydrolase involved in N-glycan trimming and is located within the Golgi, where it allows ER-escaped glycoproteins to bypass the classical N-glycosylation trimming pathway involving ER glucosidases I and II. There is considerable interest in the use of small molecules that disrupt N-linked glycosylation as therapeutic agents for diseases such as cancer and viral infection. Here we report the structure of the catalytic domain of human MANEA and complexes with substrate-derived inhibitors, which provide insight into dynamic loop movements that occur on substrate binding. We reveal structural features of the human enzyme that explain its substrate preference and the mechanistic basis for catalysis. These structures have inspired the development of new inhibitors that disrupt host protein N-glycan processing of viral glycans and reduce the infectivity of bovine viral diarrhea and dengue viruses in cellular models. These results may contribute to efforts aimed at developing broad-spectrum antiviral agents and help provide a more in-depth understanding of the biology of mammalian glycosylation.

AB - Mammalian protein N-linked glycosylation is critical for glycoprotein folding, quality control, trafficking, recognition, and function. N-linked glycans are synthesized from Glc3Man9GlcNAc2precursors that are trimmed and modified in the endoplasmic reticulum (ER) and Golgi apparatus by glycoside hydrolases and glycosyltransferases. Endo-a-1,2-mannosidase (MANEA) is the sole endoacting glycoside hydrolase involved in N-glycan trimming and is located within the Golgi, where it allows ER-escaped glycoproteins to bypass the classical N-glycosylation trimming pathway involving ER glucosidases I and II. There is considerable interest in the use of small molecules that disrupt N-linked glycosylation as therapeutic agents for diseases such as cancer and viral infection. Here we report the structure of the catalytic domain of human MANEA and complexes with substrate-derived inhibitors, which provide insight into dynamic loop movements that occur on substrate binding. We reveal structural features of the human enzyme that explain its substrate preference and the mechanistic basis for catalysis. These structures have inspired the development of new inhibitors that disrupt host protein N-glycan processing of viral glycans and reduce the infectivity of bovine viral diarrhea and dengue viruses in cellular models. These results may contribute to efforts aimed at developing broad-spectrum antiviral agents and help provide a more in-depth understanding of the biology of mammalian glycosylation.

KW - Antiviral

KW - Enzyme

KW - Glycosylation

KW - Secretory pathway

KW - Structural biology

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U2 - 10.1073/pnas.2013620117

DO - 10.1073/pnas.2013620117

M3 - Article

C2 - 33154157

AN - SCOPUS:85096884538

SN - 0027-8424

VL - 117

SP - 29595

EP - 29601

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 47

ER -

Structure of human endo-a-1,2-mannosidase (MANEA), an antiviral host-glycosylation target

Abstract

Bibliographical note

Keywords

Access to Document

Other files and links

Diffraction images used to solve the structures published in the article "Structure of human endo-α-1,2-mannosidase (MANEA), an antiviral host-glycosylation target"

Cite this

Structure of human endo-a-1,2-mannosidase (MANEA), an antiviral host-glycosylation target

Abstract

Bibliographical note

Keywords

Access to Document

Other files and links

Datasets

Diffraction images used to solve the structures published in the article "Structure of human endo-α-1,2-mannosidase (MANEA), an antiviral host-glycosylation target"

Cite this