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).

			Author Information
			(1) Heinrich Pette Institute, Leibniz Institute for
			Experimental Virology; European XFEL GmbH,
			Hamburg, Germany.
			(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/BCJ20200029 PMID: 32083638

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