tetano
Editor, Senior Moderator
Proteomics
. 2020 Oct 28;e2000246.
doi: 10.1002/pmic.202000246. Online ahead of print.
N-Terminomics for the Identification of in vitro Substrates and Cleavage Site Specificity of the SARS-CoV-2 Main Protease
Tomas Koudelka[SUP] 1 [/SUP], Juliane Boger[SUP] 2 [/SUP], Alessandra Henkel[SUP] 2 [/SUP], Robert Sch?nherr[SUP] 2 3 [/SUP], Stefanie Krantz[SUP] 4 [/SUP], Sabine Fuchs[SUP] 4 [/SUP], Estefan?a Rodr?guez[SUP] 5 [/SUP], Lars Redecke[SUP] 2 3 [/SUP], Andreas Tholey[SUP] 1 [/SUP]
Affiliations
Abstract
The genome of coronaviruses, including SARS-CoV-2, encodes for two proteases, a papain like (PL) protease and the so-called main protease (M[SUP]pro[/SUP] ), a chymotrypsin-like cysteine protease, also named 3CL[SUP]pro[/SUP] or non-structural protein 5 (nsp5). M[SUP]pro[/SUP] is activated by autoproteolysis, and is the main protease responsible for cutting the viral polyprotein into functional units. Aside from this, it has been described that M[SUP]pro[/SUP] 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, we performed an LC-MS based N-terminomics in vitro analysis using recombinantly expressed proteases and lung epithelial and endothelial cell lysates as substrate pools. For SARS-CoV-2 M[SUP]pro[/SUP] we identified 445 cleavage events from over 300 proteins, while 151 and 331 M[SUP]pro[/SUP] derived cleavage events were 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. This article is protected by copyright. All rights reserved.
Keywords: Covid-19; LC-MS; isobaric labeling; protease substrates; terminomics.
. 2020 Oct 28;e2000246.
doi: 10.1002/pmic.202000246. Online ahead of print.
N-Terminomics for the Identification of in vitro Substrates and Cleavage Site Specificity of the SARS-CoV-2 Main Protease
Tomas Koudelka[SUP] 1 [/SUP], Juliane Boger[SUP] 2 [/SUP], Alessandra Henkel[SUP] 2 [/SUP], Robert Sch?nherr[SUP] 2 3 [/SUP], Stefanie Krantz[SUP] 4 [/SUP], Sabine Fuchs[SUP] 4 [/SUP], Estefan?a Rodr?guez[SUP] 5 [/SUP], Lars Redecke[SUP] 2 3 [/SUP], Andreas Tholey[SUP] 1 [/SUP]
Affiliations
- PMID: 33111431
- DOI: 10.1002/pmic.202000246
Abstract
The genome of coronaviruses, including SARS-CoV-2, encodes for two proteases, a papain like (PL) protease and the so-called main protease (M[SUP]pro[/SUP] ), a chymotrypsin-like cysteine protease, also named 3CL[SUP]pro[/SUP] or non-structural protein 5 (nsp5). M[SUP]pro[/SUP] is activated by autoproteolysis, and is the main protease responsible for cutting the viral polyprotein into functional units. Aside from this, it has been described that M[SUP]pro[/SUP] 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, we performed an LC-MS based N-terminomics in vitro analysis using recombinantly expressed proteases and lung epithelial and endothelial cell lysates as substrate pools. For SARS-CoV-2 M[SUP]pro[/SUP] we identified 445 cleavage events from over 300 proteins, while 151 and 331 M[SUP]pro[/SUP] derived cleavage events were 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. This article is protected by copyright. All rights reserved.
Keywords: Covid-19; LC-MS; isobaric labeling; protease substrates; terminomics.