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...
| Main Authors: | , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2023-05-01
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| Series: | Frontiers in Bioengineering and Biotechnology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1180044/full |
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| author | Shiva Izadi Ulrike Vavra Stanislav Melnik Clemens Grünwald-Gruber Esther Föderl-Höbenreich Markus Sack Kurt Zatloukal Josef Glössl Eva Stöger Lukas Mach Alexandra Castilho Richard Strasser |
| author_facet | Shiva Izadi Ulrike Vavra Stanislav Melnik Clemens Grünwald-Gruber Esther Föderl-Höbenreich Markus Sack Kurt Zatloukal Josef Glössl Eva Stöger Lukas Mach Alexandra Castilho Richard Strasser |
| author_sort | Shiva Izadi |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-09T14:37:11Z |
| format | Article |
| id | doaj.art-b00aae2617684bc2bb46c3d926f815b9 |
| institution | Directory Open Access Journal |
| issn | 2296-4185 |
| language | English |
| last_indexed | 2024-04-09T14:37:11Z |
| publishDate | 2023-05-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Bioengineering and Biotechnology |
| spelling | doaj.art-b00aae2617684bc2bb46c3d926f815b92023-05-03T12:26:56ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852023-05-011110.3389/fbioe.2023.11800441180044In planta deglycosylation improves the SARS-CoV-2 neutralization activity of recombinant ACE2-FcShiva Izadi0Ulrike Vavra1Stanislav Melnik2Clemens Grünwald-Gruber3Esther Föderl-Höbenreich4Markus Sack5Kurt Zatloukal6Josef Glössl7Eva Stöger8Lukas Mach9Alexandra Castilho10Richard Strasser11Institute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences Vienna, Vienna, AustriaInstitute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences Vienna, Vienna, AustriaInstitute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences Vienna, Vienna, AustriaCore Facility Mass Spectrometry, University of Natural Resources and Life Sciences Vienna, Vienna, AustriaDiagnostic and Research Institute of Pathology, Medical University of Graz, Graz, AustriaPro-SPR GmbH, Alsdorf, GermanyDiagnostic and Research Institute of Pathology, Medical University of Graz, Graz, AustriaInstitute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences Vienna, Vienna, AustriaInstitute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences Vienna, Vienna, AustriaInstitute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences Vienna, Vienna, AustriaInstitute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences Vienna, Vienna, AustriaInstitute of Plant Biotechnology and Cell Biology, Department of Applied Genetics and Cell Biology, University of Natural Resources and Life Sciences Vienna, Vienna, AustriaSARS-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.https://www.frontiersin.org/articles/10.3389/fbioe.2023.1180044/fullCOVID-19glycosylationposttranslational modificationrecombinant protein expressionglycoengineering |
| spellingShingle | Shiva Izadi Ulrike Vavra Stanislav Melnik Clemens Grünwald-Gruber Esther Föderl-Höbenreich Markus Sack Kurt Zatloukal Josef Glössl Eva Stöger Lukas Mach Alexandra Castilho Richard Strasser In planta deglycosylation improves the SARS-CoV-2 neutralization activity of recombinant ACE2-Fc Frontiers in Bioengineering and Biotechnology COVID-19 glycosylation posttranslational modification recombinant protein expression glycoengineering |
| 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 | COVID-19 glycosylation posttranslational modification recombinant protein expression glycoengineering |
| url | https://www.frontiersin.org/articles/10.3389/fbioe.2023.1180044/full |
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