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Biophysical and structural characterizations of the effects of mutations on the structure–activity relationships of SARS-CoV-2 spike protein
Mutations on the spike (S) protein of SARS-CoV-2 could induce structural changes that help increase viral transmissibility and enhance resistance to antibody neutralization. Here, we report a robust workflow to prepare recombinant S protein variants and its host receptor angiotensin-convert enzyme 2...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier Inc.
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9388326/ https://www.ncbi.nlm.nih.gov/pubmed/36220274 http://dx.doi.org/10.1016/bs.mie.2022.07.013 |
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author | Yu, Pei-Yu Yang, Tzu-Jing Hsu, Shang-Te Danny |
author_facet | Yu, Pei-Yu Yang, Tzu-Jing Hsu, Shang-Te Danny |
author_sort | Yu, Pei-Yu |
collection | PubMed |
description | Mutations on the spike (S) protein of SARS-CoV-2 could induce structural changes that help increase viral transmissibility and enhance resistance to antibody neutralization. Here, we report a robust workflow to prepare recombinant S protein variants and its host receptor angiotensin-convert enzyme 2 (ACE2) by using a mammalian cell expression system. The functional states of the S protein variants are investigated by cryo-electron microscopy (cryo-EM) and negative staining electron microscopy (NSEM) to visualize their molecular structures in response to mutations, receptor binding, antibody binding, and environmental changes. The folding stabilities of the S protein variants can be deduced from morphological changes based on NSEM imaging analysis. Differential scanning calorimetry provides thermodynamic information to complement NSEM. Impacts of the mutations on host receptor binding and antibody neutralization are in vitro by kinetic binding analyses in addition to atomic insights gleaned from cryo-electron microscopy (cryo-EM). This experimental strategy is generally applicable to studying the molecular basis of host-pathogen interactions. |
format | Online Article Text |
id | pubmed-9388326 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Elsevier Inc. |
record_format | MEDLINE/PubMed |
spelling | pubmed-93883262022-08-19 Biophysical and structural characterizations of the effects of mutations on the structure–activity relationships of SARS-CoV-2 spike protein Yu, Pei-Yu Yang, Tzu-Jing Hsu, Shang-Te Danny Methods Enzymol Article Mutations on the spike (S) protein of SARS-CoV-2 could induce structural changes that help increase viral transmissibility and enhance resistance to antibody neutralization. Here, we report a robust workflow to prepare recombinant S protein variants and its host receptor angiotensin-convert enzyme 2 (ACE2) by using a mammalian cell expression system. The functional states of the S protein variants are investigated by cryo-electron microscopy (cryo-EM) and negative staining electron microscopy (NSEM) to visualize their molecular structures in response to mutations, receptor binding, antibody binding, and environmental changes. The folding stabilities of the S protein variants can be deduced from morphological changes based on NSEM imaging analysis. Differential scanning calorimetry provides thermodynamic information to complement NSEM. Impacts of the mutations on host receptor binding and antibody neutralization are in vitro by kinetic binding analyses in addition to atomic insights gleaned from cryo-electron microscopy (cryo-EM). This experimental strategy is generally applicable to studying the molecular basis of host-pathogen interactions. Elsevier Inc. 2022 2022-08-19 /pmc/articles/PMC9388326/ /pubmed/36220274 http://dx.doi.org/10.1016/bs.mie.2022.07.013 Text en Copyright © 2022 Elsevier Inc. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Article Yu, Pei-Yu Yang, Tzu-Jing Hsu, Shang-Te Danny Biophysical and structural characterizations of the effects of mutations on the structure–activity relationships of SARS-CoV-2 spike protein |
title | Biophysical and structural characterizations of the effects of mutations on the structure–activity relationships of SARS-CoV-2 spike protein |
title_full | Biophysical and structural characterizations of the effects of mutations on the structure–activity relationships of SARS-CoV-2 spike protein |
title_fullStr | Biophysical and structural characterizations of the effects of mutations on the structure–activity relationships of SARS-CoV-2 spike protein |
title_full_unstemmed | Biophysical and structural characterizations of the effects of mutations on the structure–activity relationships of SARS-CoV-2 spike protein |
title_short | Biophysical and structural characterizations of the effects of mutations on the structure–activity relationships of SARS-CoV-2 spike protein |
title_sort | biophysical and structural characterizations of the effects of mutations on the structure–activity relationships of sars-cov-2 spike protein |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9388326/ https://www.ncbi.nlm.nih.gov/pubmed/36220274 http://dx.doi.org/10.1016/bs.mie.2022.07.013 |
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