دورية أكاديمية
N-Glycosylation of the SARS-CoV-2 Receptor Binding Domain Is Important for Functional Expression in Plants.
| العنوان: | N-Glycosylation of the SARS-CoV-2 Receptor Binding Domain Is Important for Functional Expression in Plants. |
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| المؤلفون: | Shin YJ; Department of Applied Genetics and Cell Biology, Institute of Plant Biotechnology and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Vienna, Austria., König-Beihammer J; Department of Applied Genetics and Cell Biology, Institute of Plant Biotechnology and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Vienna, Austria., Vavra U; Department of Applied Genetics and Cell Biology, Institute of Plant Biotechnology and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Vienna, Austria., Schwestka J; Department of Applied Genetics and Cell Biology, Institute of Plant Biotechnology and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Vienna, Austria., Kienzl NF; Department of Applied Genetics and Cell Biology, Institute of Plant Biotechnology and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Vienna, Austria., Klausberger M; Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Vienna, Austria., Laurent E; Department of Biotechnology, Core Facility Biomolecular and Cellular Analysis, University of Natural Resources and Life Sciences, Vienna, Vienna, Austria., Grünwald-Gruber C; Department of Chemistry, Core Facility Mass Spectrometry, University of Natural Resources and Life Sciences, Vienna, Vienna, Austria., Vierlinger K; Competence Unit Molecular Diagnostics, Center for Health and Bioresources, AIT Austrian Institute of Technology GmbH, Vienna, Austria., Hofner M; Competence Unit Molecular Diagnostics, Center for Health and Bioresources, AIT Austrian Institute of Technology GmbH, Vienna, Austria., Margolin E; Division of Medical Virology, Department of Pathology, Faculty of Health Sciences, University of Cape Town, Cape Town, South Africa.; Biopharming Research Unit, Department of Molecular and Cell Biology, University of Cape Town, Cape Town, South Africa., Weinhäusel A; Competence Unit Molecular Diagnostics, Center for Health and Bioresources, AIT Austrian Institute of Technology GmbH, Vienna, Austria., Stöger E; Department of Applied Genetics and Cell Biology, Institute of Plant Biotechnology and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Vienna, Austria., Mach L; Department of Applied Genetics and Cell Biology, Institute of Plant Biotechnology and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Vienna, Austria., Strasser R; Department of Applied Genetics and Cell Biology, Institute of Plant Biotechnology and Cell Biology, University of Natural Resources and Life Sciences, Vienna, Vienna, Austria. |
| المصدر: | Frontiers in plant science [Front Plant Sci] 2021 Jun 15; Vol. 12, pp. 689104. Date of Electronic Publication: 2021 Jun 15 (Print Publication: 2021). |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Frontiers Research Foundation Country of Publication: Switzerland NLM ID: 101568200 Publication Model: eCollection Cited Medium: Print ISSN: 1664-462X (Print) Linking ISSN: 1664462X NLM ISO Abbreviation: Front Plant Sci Subsets: PubMed not MEDLINE |
| أسماء مطبوعة: | Original Publication: Lausanne : Frontiers Research Foundation, 2010- |
| مستخلص: | Nicotiana benthamiana is used worldwide as production host for recombinant proteins. Many recombinant proteins such as monoclonal antibodies, growth factors or viral antigens require posttranslational modifications like glycosylation for their function. Here, we transiently expressed different variants of the glycosylated receptor binding domain (RBD) from the SARS-CoV-2 spike protein in N. benthamiana . We characterized the impact of variations in RBD-length and posttranslational modifications on protein expression, yield and functionality. We found that a truncated RBD variant (RBD-215) consisting of amino acids Arg319-Leu533 can be efficiently expressed as a secreted soluble protein. Purified RBD-215 was mainly present as a monomer and showed binding to the conformation-dependent antibody CR3022, the cellular receptor angiotensin converting enzyme 2 (ACE2) and to antibodies present in convalescent sera. Expression of RBD-215 in glycoengineered ΔXT/FT plants resulted in the generation of complex N-glycans on both N-glycosylation sites. While site-directed mutagenesis showed that the N-glycans are important for proper RBD folding, differences in N-glycan processing had no effect on protein expression and function. Competing Interests: EM is a named inventor on patent applications describing the use of chaperones to improve protein production in plants. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. (Copyright © 2021 Shin, König-Beihammer, Vavra, Schwestka, Kienzl, Klausberger, Laurent, Grünwald-Gruber, Vierlinger, Hofner, Margolin, Weinhäusel, Stöger, Mach and Strasser.) |
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| معلومات مُعتمدة: | United Kingdom WT_ Wellcome Trust; P 31920 Austria FWF_ Austrian Science Fund FWF; W 1224 Austria FWF_ Austrian Science Fund FWF |
| فهرسة مساهمة: | Keywords: COVID-19; SARS-CoV-2; glycoprotein; glycosylation; posttranslational modification; virus |
| تواريخ الأحداث: | Date Created: 20210702 Latest Revision: 20231107 |
| رمز التحديث: | 20240628 |
| مُعرف محوري في PubMed: | PMC8239413 |
| DOI: | 10.3389/fpls.2021.689104 |
| PMID: | 34211491 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1664-462X |
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| DOI: | 10.3389/fpls.2021.689104 |