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Production of Proteins of the SARS-CoV-2 Proteome for Drug Discovery

[Image: see text] The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is the causative agent of the coronavirus disease of 2019 (COVID-19). Its genome encodes two open reading frames for two large proteins, PP1a and PP1ab. Within the two polypeptide stretches, there are two proteases th...

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Autores principales: Kim, Choon, Mahasenan, Kiran V., Bhardwaj, Atul, Wiest, Olaf, Chang, Mayland, Mobashery, Shahriar
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2021
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8315141/
https://www.ncbi.nlm.nih.gov/pubmed/34337272
http://dx.doi.org/10.1021/acsomega.1c02984
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author Kim, Choon
Mahasenan, Kiran V.
Bhardwaj, Atul
Wiest, Olaf
Chang, Mayland
Mobashery, Shahriar
author_facet Kim, Choon
Mahasenan, Kiran V.
Bhardwaj, Atul
Wiest, Olaf
Chang, Mayland
Mobashery, Shahriar
author_sort Kim, Choon
collection PubMed
description [Image: see text] The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is the causative agent of the coronavirus disease of 2019 (COVID-19). Its genome encodes two open reading frames for two large proteins, PP1a and PP1ab. Within the two polypeptide stretches, there are two proteases that process the large proteins into 15 discrete proteins essential for the assembly of the virion during its replication. We describe herein the cloning of the genes for these discrete proteins optimized for expression in Escherichia coli, production of the proteins, and their purification to homogeneity. These included all but six: NSP6, which possesses eight transmembrane regions, and five that are small proteins/peptides (E, ORF3b, ORF6, ORF7b, and ORF10). These proteins are intended for experimental validation of small-molecule binders as molecular template hits. The proof of concept was established with the ADP-ribosylhydrolase (ARH) domain of NSP3 in discovery of small-molecule templates that could serve as the basis for further optimization. The hit molecules include one submicromolar and a few low-micromolar binders to the ARH domain. Availability of these proteins in soluble forms opens up the opportunity for discoveries of novel templates with the potential for anti-COVID-19 pharmaceuticals.
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spelling pubmed-83151412021-07-27 Production of Proteins of the SARS-CoV-2 Proteome for Drug Discovery Kim, Choon Mahasenan, Kiran V. Bhardwaj, Atul Wiest, Olaf Chang, Mayland Mobashery, Shahriar ACS Omega [Image: see text] The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is the causative agent of the coronavirus disease of 2019 (COVID-19). Its genome encodes two open reading frames for two large proteins, PP1a and PP1ab. Within the two polypeptide stretches, there are two proteases that process the large proteins into 15 discrete proteins essential for the assembly of the virion during its replication. We describe herein the cloning of the genes for these discrete proteins optimized for expression in Escherichia coli, production of the proteins, and their purification to homogeneity. These included all but six: NSP6, which possesses eight transmembrane regions, and five that are small proteins/peptides (E, ORF3b, ORF6, ORF7b, and ORF10). These proteins are intended for experimental validation of small-molecule binders as molecular template hits. The proof of concept was established with the ADP-ribosylhydrolase (ARH) domain of NSP3 in discovery of small-molecule templates that could serve as the basis for further optimization. The hit molecules include one submicromolar and a few low-micromolar binders to the ARH domain. Availability of these proteins in soluble forms opens up the opportunity for discoveries of novel templates with the potential for anti-COVID-19 pharmaceuticals. American Chemical Society 2021-07-20 /pmc/articles/PMC8315141/ /pubmed/34337272 http://dx.doi.org/10.1021/acsomega.1c02984 Text en © 2021 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by-nc-nd/4.0/Permits non-commercial access and re-use, provided that author attribution and integrity are maintained; but does not permit creation of adaptations or other derivative works (https://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Kim, Choon
Mahasenan, Kiran V.
Bhardwaj, Atul
Wiest, Olaf
Chang, Mayland
Mobashery, Shahriar
Production of Proteins of the SARS-CoV-2 Proteome for Drug Discovery
title Production of Proteins of the SARS-CoV-2 Proteome for Drug Discovery
title_full Production of Proteins of the SARS-CoV-2 Proteome for Drug Discovery
title_fullStr Production of Proteins of the SARS-CoV-2 Proteome for Drug Discovery
title_full_unstemmed Production of Proteins of the SARS-CoV-2 Proteome for Drug Discovery
title_short Production of Proteins of the SARS-CoV-2 Proteome for Drug Discovery
title_sort production of proteins of the sars-cov-2 proteome for drug discovery
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8315141/
https://www.ncbi.nlm.nih.gov/pubmed/34337272
http://dx.doi.org/10.1021/acsomega.1c02984
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