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In planta deglycosylation improves the SARS-CoV-2 neutralization activity of recombinant ACE2-Fc
SARS-CoV-2 infects human cells via binding of the viral spike glycoprotein to its main cellular receptor, angiotensin-converting enzyme 2 (ACE2). The spike protein-ACE2 receptor interaction is therefore a major target for the development of therapeutic or prophylactic drugs to combat coronavirus inf...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10190127/ https://www.ncbi.nlm.nih.gov/pubmed/37207124 http://dx.doi.org/10.3389/fbioe.2023.1180044 |
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author | Izadi, Shiva Vavra, Ulrike Melnik, Stanislav Grünwald-Gruber, Clemens Föderl-Höbenreich, Esther Sack, Markus Zatloukal, Kurt Glössl, Josef Stöger, Eva Mach, Lukas Castilho, Alexandra Strasser, Richard |
author_facet | Izadi, Shiva Vavra, Ulrike Melnik, Stanislav Grünwald-Gruber, Clemens Föderl-Höbenreich, Esther Sack, Markus Zatloukal, Kurt Glössl, Josef Stöger, Eva Mach, Lukas Castilho, Alexandra Strasser, Richard |
author_sort | Izadi, Shiva |
collection | PubMed |
description | SARS-CoV-2 infects human cells via binding of the viral spike glycoprotein to its main cellular receptor, angiotensin-converting enzyme 2 (ACE2). The spike protein-ACE2 receptor interaction is therefore a major target for the development of therapeutic or prophylactic drugs to combat coronavirus infections. Various engineered soluble ACE2 variants (decoys) have been designed and shown to exhibit virus neutralization capacity in cell-based assays and in vivo models. Human ACE2 is heavily glycosylated and some of its glycans impair binding to the SARS-CoV-2 spike protein. Therefore, glycan-engineered recombinant soluble ACE2 variants might display enhanced virus-neutralization potencies. Here, we transiently co-expressed the extracellular domain of ACE2 fused to human Fc (ACE2-Fc) with a bacterial endoglycosidase in Nicotiana benthamiana to produce ACE2-Fc decorated with N-glycans consisting of single GlcNAc residues. The endoglycosidase was targeted to the Golgi apparatus with the intention to avoid any interference of glycan removal with concomitant ACE2-Fc protein folding and quality control in the endoplasmic reticulum. The in vivo deglycosylated ACE2-Fc carrying single GlcNAc residues displayed increased affinity to the receptor-binding domain (RBD) of SARS-CoV-2 as well as improved virus neutralization activity and thus is a promising drug candidate to block coronavirus infection. |
format | Online Article Text |
id | pubmed-10190127 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-101901272023-05-18 In planta deglycosylation improves the SARS-CoV-2 neutralization activity of recombinant ACE2-Fc Izadi, Shiva Vavra, Ulrike Melnik, Stanislav Grünwald-Gruber, Clemens Föderl-Höbenreich, Esther Sack, Markus Zatloukal, Kurt Glössl, Josef Stöger, Eva Mach, Lukas Castilho, Alexandra Strasser, Richard Front Bioeng Biotechnol Bioengineering and Biotechnology SARS-CoV-2 infects human cells via binding of the viral spike glycoprotein to its main cellular receptor, angiotensin-converting enzyme 2 (ACE2). The spike protein-ACE2 receptor interaction is therefore a major target for the development of therapeutic or prophylactic drugs to combat coronavirus infections. Various engineered soluble ACE2 variants (decoys) have been designed and shown to exhibit virus neutralization capacity in cell-based assays and in vivo models. Human ACE2 is heavily glycosylated and some of its glycans impair binding to the SARS-CoV-2 spike protein. Therefore, glycan-engineered recombinant soluble ACE2 variants might display enhanced virus-neutralization potencies. Here, we transiently co-expressed the extracellular domain of ACE2 fused to human Fc (ACE2-Fc) with a bacterial endoglycosidase in Nicotiana benthamiana to produce ACE2-Fc decorated with N-glycans consisting of single GlcNAc residues. The endoglycosidase was targeted to the Golgi apparatus with the intention to avoid any interference of glycan removal with concomitant ACE2-Fc protein folding and quality control in the endoplasmic reticulum. The in vivo deglycosylated ACE2-Fc carrying single GlcNAc residues displayed increased affinity to the receptor-binding domain (RBD) of SARS-CoV-2 as well as improved virus neutralization activity and thus is a promising drug candidate to block coronavirus infection. Frontiers Media S.A. 2023-05-03 /pmc/articles/PMC10190127/ /pubmed/37207124 http://dx.doi.org/10.3389/fbioe.2023.1180044 Text en Copyright © 2023 Izadi, Vavra, Melnik, Grünwald-Gruber, Föderl-Höbenreich, Sack, Zatloukal, Glössl, Stöger, Mach, Castilho and Strasser. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Bioengineering and Biotechnology Izadi, Shiva Vavra, Ulrike Melnik, Stanislav Grünwald-Gruber, Clemens Föderl-Höbenreich, Esther Sack, Markus Zatloukal, Kurt Glössl, Josef Stöger, Eva Mach, Lukas Castilho, Alexandra Strasser, Richard In planta deglycosylation improves the SARS-CoV-2 neutralization activity of recombinant ACE2-Fc |
title |
In planta deglycosylation improves the SARS-CoV-2 neutralization activity of recombinant ACE2-Fc |
title_full |
In planta deglycosylation improves the SARS-CoV-2 neutralization activity of recombinant ACE2-Fc |
title_fullStr |
In planta deglycosylation improves the SARS-CoV-2 neutralization activity of recombinant ACE2-Fc |
title_full_unstemmed |
In planta deglycosylation improves the SARS-CoV-2 neutralization activity of recombinant ACE2-Fc |
title_short |
In planta deglycosylation improves the SARS-CoV-2 neutralization activity of recombinant ACE2-Fc |
title_sort | in planta deglycosylation improves the sars-cov-2 neutralization activity of recombinant ace2-fc |
topic | Bioengineering and Biotechnology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10190127/ https://www.ncbi.nlm.nih.gov/pubmed/37207124 http://dx.doi.org/10.3389/fbioe.2023.1180044 |
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