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PXD049009-1

PXD049009 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleThe kinetics of nsp7-11 polyprotein processing and impact on complexation with nsp16 among human coronaviruses
DescriptionIn coronavirus (CoV) infection, polyproteins (pp1a/pp1ab) are processed into non-structural proteins (nsps), which largely form the replication/transcription complex (RTC). The polyprotein processing and complex formation is critical and offers potential therapeutic targets. However, the interplay of polyprotein processing and RTC-assembly remains poorly understood. Here, we studied two key aspects: The order of polyprotein processing by viral main protease Mpro and its influence on complex formation with the methyltransferase nsp16. Moreover, we established an approach to determine rate constants k from cleavage sites in structured CoV polyprotein based on native mass spectrometry (MS). The high sensitivity and precision of our method allowed quantification of multi-reaction kinetics of nsp7-11 processing from four human pathogenic CoV species. The experimentally determined rate constants are put into perspective with a comprehensive analysis of primary sequences and structural models, revealing distinct cleavage mechanisms for each site based on their local structural environments. Our systematic approach provides a blueprint for kinetic analysis of complex multi-cleavage reactions.
HostingRepositoryPRIDE
AnnounceDate2025-07-08
AnnouncementXMLSubmission_2025-07-08_14:24:41.610.xml
DigitalObjectIdentifier
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportUnsupported dataset by repository
PrimarySubmitterKira Schamoni-Kast
SpeciesList scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationListNo PTMs are included in the dataset
InstrumentQ-Tof Ultima; Q Exactive
Dataset History
RevisionDatetimeStatusChangeLog Entry
02024-01-30 06:51:41ID requested
12025-07-08 14:24:41announced
Publication List
10.1038/S41467-025-61554-Y;
Keyword List
submitter keyword: mass spectrometry, SARS-CoV-2,Kinetics, native MS, rate constants, complexation
Contact List
Charlotte Uetrecht
contact affiliationCSSB Centre for Structural Systems Biology, Deutsches Elektronen Synchroton DESY, Leibniz Institute of Virology, Germany; University of Luebeck, Institute for Chemistry and Metabolomics
contact emailcharlotte.uetrecht@cssb-hamburg.de
lab head
Kira Schamoni-Kast
contact affiliationCentre for Structural Systems Biology (CSSB), Leibniz Institute of Virology (LIV)
contact emailkira.schamoni-kast@cssb-hamburg.de
dataset submitter
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Dataset FTP location
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