PubMed Journals: Biochem J
Source: PMID: 32083638⇦⇨ Biochem J. 2020 Feb 21. pii: BCJ20200029.
⇩ doi: 10.1042/BCJ20200029. [Epub ahead of
Processing of the SARS-CoV pp1a/ab nsp7-10
Krichel B(1), Falke S(2), Hilgenfeld R(3),
Redecke L(4), Uetrecht C(1).
(1) Heinrich Pette Institute, Leibniz Institute for
Experimental Virology; European XFEL GmbH,
(2) Universität Hamburg, Hamburg, Germany.
(3) University of Lübeck, Lübeck, Germany.
(4) University of Lübeck.
Severe acute respiratory syndrome coronavirus
(SARS-CoV) is the causative agent of a
respiratory disease with a high case fatality
rate. During the formation of the coronaviral
replication/transcription complex (RTC),
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
Copyright 2020 The Author(s).
DOI: 10.1042/BCJ20200029PMID: 32083638⇧TweetPrint