biostudies-literatureVlok MCanadian Government | Canadian Institutes of Health ResearchDST | Science and Engineering Research BoardDST | Science and Engineering Research Board (SERB)Canadian Government | Natural Sciences and Engineering Research Council of Canada (NSERC)Canadian Government | Natural Sciences and Engineering Research Council of CanadaCanadian Government | Canadian Institutes of Health Research (CIHR)e0049824https://www.ebi.ac.uk/biostudies/studies/S-EPMC11265341biostudies-literatureUnknown98(7)Coxsackievirus B3 (CVB3) encodes proteinases that are essential for processing of the translated viral polyprotein. Viral proteinases also target host proteins to manipulate cellular processes and evade innate antiviral responses to promote replication and infection. While some host protein substrates of the CVB3 3C and 2A cysteine proteinases have been identified, the full repertoire of targets is not known. Here, we utilize an unbiased quantitative proteomics-based approach termed terminal amine isotopic labeling of substrates (TAILS) to conduct a global analysis of CVB3 protease-generated N-terminal peptides in both human HeLa and mouse cardiomyocyte (HL-1) cell lines infected with CVB3. We identified >800 proteins that are cleaved in CVB3-infected HeLa and HL-1 cells including the viraJournal of virologyIdentification of the proteolytic signature in CVB3-infected cells.PMC11265341PJT-156351, PJT-173318, FDN-148408RGPIN-2017-04515, RGPIN-2022-02979, PGS DSadasivan JMohamud YOverall CMVlok MSolis NWarsaba RLuo HJan EKizhakkedathu JfalseIdentification of the proteolytic signature in CVB3-infected cells.Coxsackievirus B3 (CVB3) encodes proteinases that are essential for processing of the translated viral polyprotein. Viral proteinases also target host proteins to manipulate cellular processes and evade innate antiviral responses to promote replication and infection. While some host protein substrates of the CVB3 3C and 2A cysteine proteinases have been identified, the full repertoire of targets is not known. Here, we utilize an unbiased quantitative proteomics-based approach termed terminal amine isotopic labeling of substrates (TAILS) to conduct a global analysis of CVB3 protease-generated N-terminal peptides in both human HeLa and mouse cardiomyocyte (HL-1) cell lines infected with CVB3. We identified >800 proteins that are cleaved in CVB3-infected HeLa and HL-1 cells including the vira2024-01-01T00:00:00Z2024 Jul2026-06-02T05:03:29.722Z2025-04-19T13:13:52.94ZS-EPMC112653413895366710.1128/jvi.00498-24