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dc.contributor.authorSandu, Cristinelen_US
dc.contributor.authorChandramouli, Nagaranjanen_US
dc.contributor.authorGlickman, Joseph Fraseren_US
dc.contributor.authorMolina, Henriken_US
dc.contributor.authorKuo, Chueh-Lingen_US
dc.contributor.authorKukushkin, Nikolayen_US
dc.contributor.authorGoldberg, Alfred Len_US
dc.contributor.authorSteller, Hermannen_US
dc.date.accessioned2015-10-01T14:57:14Z
dc.date.issued2015en_US
dc.identifier.citationSandu, Cristinel, Nagaranjan Chandramouli, Joseph Fraser Glickman, Henrik Molina, Chueh-Ling Kuo, Nikolay Kukushkin, Alfred L Goldberg, and Hermann Steller. 2015. “Thiostrepton interacts covalently with Rpt subunits of the 19S proteasome and proteasome substrates.” Journal of Cellular and Molecular Medicine 19 (9): 2181-2192. doi:10.1111/jcmm.12602. http://dx.doi.org/10.1111/jcmm.12602.en
dc.identifier.issn1582-1838en
dc.identifier.urihttp://nrs.harvard.edu/urn-3:HUL.InstRepos:22856985
dc.description.abstractHere, we report a novel mechanism of proteasome inhibition mediated by Thiostrepton (Thsp), which interacts covalently with Rpt subunits of the 19S proteasome and proteasome substrates. We identified Thsp in a cell-based high-throughput screen using a fluorescent reporter sensitive to degradation by the ubiquitin–proteasome pathway. Thiostrepton behaves as a proteasome inhibitor in several paradigms, including cell-based reporters, detection of global ubiquitination status, and proteasome-mediated labile protein degradation. In vitro, Thsp does not block the chymotrypsin activity of the 26S proteasome. In a cell-based IκBα degradation assay, Thsp is a slow inhibitor and 4 hrs of treatment achieves the same effects as MG-132 at 30 min. We show that Thsp forms covalent adducts with proteins in human cells and demonstrate their nature by mass spectrometry. Furthermore, the ability of Thsp to interact covalently with the cysteine residues is essential for its proteasome inhibitory function. We further show that a Thsp modified peptide cannot be degraded by proteasomes in vitro. Importantly, we demonstrate that Thsp binds covalently to Rpt subunits of the 19S regulatory particle and forms bridges with a proteasome substrate. Taken together, our results uncover an important role of Thsp in 19S proteasome inhibition.en
dc.language.isoen_USen
dc.publisherJohn Wiley & Sons, Ltden
dc.relation.isversionofdoi:10.1111/jcmm.12602en
dc.relation.hasversionhttp://www.ncbi.nlm.nih.gov/pmc/articles/PMC4568923/pdf/en
dash.licenseLAAen_US
dc.subjectthiolen
dc.subjectprotein degradationen
dc.subjectprotein chemical modificationen
dc.subjectproteasomeen
dc.subjectubiquitinen
dc.titleThiostrepton interacts covalently with Rpt subunits of the 19S proteasome and proteasome substratesen
dc.typeJournal Articleen_US
dc.description.versionVersion of Recorden
dc.relation.journalJournal of Cellular and Molecular Medicineen
dash.depositing.authorKuo, Chueh-Lingen_US
dc.date.available2015-10-01T14:57:14Z
dc.identifier.doi10.1111/jcmm.12602*
dash.contributor.affiliatedKuo, Chueh-Ling
dash.contributor.affiliatedGoldberg, Alfred


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