TY - JOUR
T1 - Processing of the SARS-CoV pp1a/ab nsp7-10 region
AU - Krichel, Boris
AU - Falke, Sven
AU - Hilgenfeld, Rolf
AU - Redecke, Lars
AU - Uetrecht, Charlotte
N1 - Funding Information:
The Heinrich Pette Institute, Leibniz Institute for Experimental Virology is supported by the Free and Hanseatic City of Hamburg and the Federal Ministry of Health. C.U. and B.K. are supported by EU Horizon 2020 ERC StG-2017 759661. C.U. and B.K. also received funding through the Leibniz Association SAW-2014-HPI-4 grant. S.F. and L.R. thank the German Federal Ministry for Education and Research (BMBF) for funding [grant numbers 01KX0806, 01KX0807 and 05K18FLA]. R.H. acknowledges funding from the SILVER project of the European Commission (contract HEALTH-F3-2010-260644).
Publisher Copyright:
© 2020 The Author(s). This is an open access article published by Portland Press Limited on behalf of the Biochemical Society and distributed under the Creative Commons Attribution License 4.0 (CC BY).
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2020/3/13
Y1 - 2020/3/13
N2 - Severe acute respiratory syndrome coronavirus is the causative agent of a respiratory disease with a high case fatality rate. During the formation of the coronaviral replication/ transcription complex, essential steps include processing of the conserved polyprotein nsp7-10 region by the main protease Mpro and subsequent complex formation of the released nsp's. Here, we analyzed processing of the coronavirus nsp7-10 region using native mass spectrometry showing consumption of substrate, rise and fall of intermediate products and complexation. Importantly, there is a clear order of cleavage efficiencies, which is influenced by the polyprotein tertiary structure. Furthermore, the predominant product is an nsp7+8(2: 2) hetero-tetramer with nsp8 scaffold. In conclusion, native MS, opposed to other methods, can expose the processing dynamics of viral polyproteins and the landscape of protein interactions in one set of experiments. Thereby, new insights into protein interactions, essential for generation of viral progeny, were provided, with relevance for development of antivirals.
AB - Severe acute respiratory syndrome coronavirus is the causative agent of a respiratory disease with a high case fatality rate. During the formation of the coronaviral replication/ transcription complex, essential steps include processing of the conserved polyprotein nsp7-10 region by the main protease Mpro and subsequent complex formation of the released nsp's. Here, we analyzed processing of the coronavirus nsp7-10 region using native mass spectrometry showing consumption of substrate, rise and fall of intermediate products and complexation. Importantly, there is a clear order of cleavage efficiencies, which is influenced by the polyprotein tertiary structure. Furthermore, the predominant product is an nsp7+8(2: 2) hetero-tetramer with nsp8 scaffold. In conclusion, native MS, opposed to other methods, can expose the processing dynamics of viral polyproteins and the landscape of protein interactions in one set of experiments. Thereby, new insights into protein interactions, essential for generation of viral progeny, were provided, with relevance for development of antivirals.
UR - http://www.scopus.com/inward/record.url?scp=85082146457&partnerID=8YFLogxK
U2 - 10.1042/BCJ20200029
DO - 10.1042/BCJ20200029
M3 - Journal articles
C2 - 32083638
AN - SCOPUS:85082146457
SN - 0264-6021
VL - 477
SP - 1009
EP - 1019
JO - Biochemical Journal
JF - Biochemical Journal
IS - 5
ER -
