Dataset Information

N-terminomics for the identification of in-vitro substrates and cleavage site specificity of the SARS-CoV-2 main protease

ABSTRACT: The genome of coronaviruses, including the SARS-CoV-2, which causes the human pandemic COVID-19, encodes for two proteases, a papain like (PL) and the so-called Main protease (Mpro), also named 3CLpro or non-structural protein 5 (NSP5). Mpro is activated by autoproteolysis, and true to its name, is the main protease responsible for cutting the viral polyprotein into functional viral proteins. Aside from this function, it has been described that 3CL proteases are also capable to process host proteins, including those involved in the host innate immune response. We performed a LC-MS based N-terminomics analysis to identify in vitro substrates of three different recombinantly expressed coronavirus main proteases (SARS-CoV, SARS-CoV-2, hCoV-NL63) using lung cell lysates as substrate pools.

INSTRUMENT(S): Orbitrap Fusion Lumos, Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

TISSUE(S): Lung

SUBMITTER: Andreas Tholey

LAB HEAD: Andreas Tholey

PROVIDER: PXD021406 | Pride | 2020-11-02

REPOSITORIES: Pride

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Publications

N-Terminomics for the Identification of In Vitro Substrates and Cleavage Site Specificity of the SARS-CoV-2 Main Protease.

Koudelka Tomas T Boger Juliane J Henkel Alessandra A Schönherr Robert R Krantz Stefanie S Fuchs Sabine S Rodríguez Estefanía E Redecke Lars L Tholey Andreas A

Proteomics 20201117 2

The genome of coronaviruses, including SARS-CoV-2, encodes for two proteases, a papain like (PLpro ) protease and the so-called main protease (Mpro ), a chymotrypsin-like cysteine protease, also named 3CLpro or non-structural protein 5 (nsp5). Mpro is activated by autoproteolysis and is the main protease responsible for cutting the viral polyprotein into functional units. Aside from this, it is described that Mpro proteases are also capable ...[more]

PMID: 33111431

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Project description:Despite the wide availability of several safe and effective vaccines that can prevent severe COVID-19 disease, the emergence of SARS-CoV-2 variants of concern (VOC) that can partially evade vaccine immunity remains a global health concern. In addition, the emergence of highly mutated and neutralization-resistant SARS-CoV-2 VOCs such as BA.1 and BA.5 that can partially or fully evade (1) many therapeutic monoclonal antibodies in clinical use underlines the need for additional effective treatment strategies. Here, we characterize the antiviral activity of GS-5245, Obeldesivir (ODV), an oral prodrug of the parent nucleoside GS-441524, which targets the highly conserved RNA-dependent viral RNA polymerase (RdRp). Importantly, we show that GS-5245 is broadly potent in vitro against alphacoronavirus HCoV-NL63, severe acute respiratory syndrome coronavirus (SARS-CoV), SARS-CoV-related Bat-CoV RsSHC014, Middle East Respiratory Syndrome coronavirus (MERS-CoV), SARS-CoV-2 WA/1, and the highly transmissible SARS-CoV-2 BA.1 Omicron variant in vitro and highly effective as antiviral therapy in mouse models of SARS-CoV, SARS-CoV-2 (WA/1), MERS-CoV and Bat-CoV RsSHC014 pathogenesis. In all these models of divergent coronaviruses, we observed protection and/or significant reduction of disease metrics such as weight loss, lung viral replication, acute lung injury, and degradation in pulmonary function in GS-5245-treated mice compared to vehicle controls. Finally, we demonstrate that GS-5245 in combination with the main protease (Mpro) inhibitor nirmatrelvir had increased efficacy in vivo against SARS-CoV-2 compared to each single agent. We also evalulate the effect of antiviral therapy on host gene expression using RNAseq and show that therapeutic intervention reduces host inflammatory response as compared to vehicle controls during acute SARS-CoV-2 infection. Altogether, our data supports the continuing clinical evaluation of GS-5245 in humans infected with COVID-19, including as part of a combination antiviral therapy, especially in populations with the most urgent need for more efficacious and durable interventions.

Dataset Information

N-terminomics for the identification of in-vitro substrates and cleavage site specificity of the SARS-CoV-2 main protease

Publications

N-Terminomics for the Identification of In Vitro Substrates and Cleavage Site Specificity of the SARS-CoV-2 Main Protease.

Similar Datasets

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