Abstract
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 of processing host proteins, including those involved in the host innate immune response. To identify substrates of the three main proteases from SARS-CoV, SARS-CoV-2, and hCoV-NL63 coronviruses, an LC-MS based N-terminomics in vitro analysis is performed using recombinantly expressed proteases and lung epithelial and endothelial cell lysates as substrate pools. For SARS-CoV-2 Mpro, 445 cleavage events from more than 300 proteins are identified, while 151 and 331 Mpro derived cleavage events are identified for SARS-CoV and hCoV-NL63, respectively. These data enable to better understand the cleavage site specificity of the viral proteases and will help to identify novel substrates in vivo. All data are available via ProteomeXchange with identifier PXD021406.
| Original language | English |
|---|---|
| Article number | 2000246 |
| Journal | Proteomics |
| Volume | 21 |
| Issue number | 2 |
| Pages (from-to) | e2000246 |
| ISSN | 1615-9853 |
| DOIs | |
| Publication status | Published - 01.2021 |
Funding
This project was funded by the Deutsche Forschungsgemeinschaft (DFG), SFB877 (project Z2), and the Cluster of Excellence “Precision Medicine in Inflammation” (PMI, RTF‐V). J.B. and L.R. thank the German Federal Ministry for Education and Research (BMBF) for funding (grant number 05K18FLA). The authors thank Fanlu Wang for technical support with the cell cultures, Rolf Hilgenfeld for providing the gene of SARS‐CoV M, and Linlin Zhang for SARS‐CoV‐2 Mpro gene and hCoV‐NL63 Mpro protein. pro This project was funded by the Deutsche Forschungsgemeinschaft (DFG), SFB877 (project Z2), and the Cluster of Excellence ?Precision Medicine in Inflammation? (PMI, RTF-V). J.B. and L.R. thank the German Federal Ministry for Education and Research (BMBF) for funding (grant number 05K18FLA). The authors thank Fanlu Wang for technical support with the cell cultures, Rolf Hilgenfeld for providing the gene of SARS-CoV Mpro, and Linlin Zhang for SARS-CoV-2 Mpro gene and hCoV-NL63 Mpro protein. Open access funding enabled and organized by Projekt DEAL.
Research Areas and Centers
- Academic Focus: Center for Infection and Inflammation Research (ZIEL)
Coronavirus related work
- Research on SARS-CoV-2 / COVID-19
