{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,23]],"date-time":"2026-07-23T02:35:43Z","timestamp":1784774143716,"version":"3.55.0"},"reference-count":48,"publisher":"Portland Press Ltd.","issue":"3","license":[{"start":{"date-parts":[[2019,7,25]],"date-time":"2019-07-25T00:00:00Z","timestamp":1564012800000},"content-version":"vor","delay-in-days":1602,"URL":"http:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":["portlandpress.com"],"crossmark-restriction":true},"short-container-title":[],"published-print":{"date-parts":[[2015,3,15]]},"abstract":"<jats:p>Modification of proteins with ubiquitin (Ub) occurs through a variety of topologically distinct Ub linkages, including Ube2W-mediated monoubiquitylation of N-terminal alpha amines to generate peptide-linked linear mono-Ub fusions. Protein ubiquitylation can be reversed by the action of deubiquitylating enzymes (DUBs), many of which show striking preference for particular Ub linkage types. Here, we have screened for DUBs that preferentially cleave N-terminal Ub from protein substrates but do not act on Ub homopolymers. We show that members of the Ub C-terminal hydrolase (UCH) family of DUBs demonstrate this preference for N-terminal deubiquitylating activity as they are capable of cleaving N-terminal Ub from SUMO2 and Ube2W, while displaying no activity against any of the eight Ub linkage types. Surprisingly, this ability to cleave Ub from SUMO2 was 100\u00a0times more efficient for UCH-L3 when we deleted the unstructured N-terminus of SUMO2, demonstrating that UCH enzymes can cleave Ub from structured proteins. However, UCH-L3 could also cleave chemically synthesized isopeptide-linked Ub from lysine 11 (K11) of SUMO2 with similar efficiency, demonstrating that UCH DUB activity is not limited to peptide-linked Ub. These findings advance our understanding of the specificity of the UCH family of DUBs, which are strongly implicated in cancer and neurodegeneration but whose substrate preference has remained unclear. In addition, our findings suggest that the reversal of Ube2W-mediated N-terminal ubiquitylation may be one physiological role of UCH DUBs in\u00a0vivo.<\/jats:p>","DOI":"10.1042\/bj20141349","type":"journal-article","created":{"date-parts":[[2014,12,15]],"date-time":"2014-12-15T04:38:37Z","timestamp":1418618317000},"page":"489-498","update-policy":"https:\/\/doi.org\/10.1042\/crossmark_policy","source":"Crossref","is-referenced-by-count":54,"title":["Ubiquitin C-terminal hydrolases cleave isopeptide- and peptide-linked ubiquitin from structured proteins but do not edit ubiquitin homopolymers"],"prefix":"10.1042","volume":"466","author":[{"given":"John\u00a0S.","family":"Bett","sequence":"first","affiliation":[{"name":"MRC Protein Phosphorylation and Ubiquitylation Unit, The Sir James Black Centre, College of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maria\u00a0Stella","family":"Ritorto","sequence":"additional","affiliation":[{"name":"MRC Protein Phosphorylation and Ubiquitylation Unit, The Sir James Black Centre, College of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Richard","family":"Ewan","sequence":"additional","affiliation":[{"name":"MRC Protein Phosphorylation and Ubiquitylation Unit, The Sir James Black Centre, College of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ellis\u00a0G.","family":"Jaffray","sequence":"additional","affiliation":[{"name":"Centre for Gene Regulation and Expression, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Satpal","family":"Virdee","sequence":"additional","affiliation":[{"name":"MRC Protein Phosphorylation and Ubiquitylation Unit, The Sir James Black Centre, College of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jason\u00a0W.","family":"Chin","sequence":"additional","affiliation":[{"name":"MRC Laboratory of Molecular Biology, Cambridge Biomedical Campus, Cambridge CB2 0QH, England, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Axel","family":"Knebel","sequence":"additional","affiliation":[{"name":"MRC Protein Phosphorylation and Ubiquitylation Unit, The Sir James Black Centre, College of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thimo","family":"Kurz","sequence":"additional","affiliation":[{"name":"MRC Protein Phosphorylation and Ubiquitylation Unit, The Sir James Black Centre, College of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Matthias","family":"Trost","sequence":"additional","affiliation":[{"name":"MRC Protein Phosphorylation and Ubiquitylation Unit, The Sir James Black Centre, College of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael\u00a0H.","family":"Tatham","sequence":"additional","affiliation":[{"name":"Centre for Gene Regulation and Expression, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ronald\u00a0T.","family":"Hay","sequence":"additional","affiliation":[{"name":"MRC Protein Phosphorylation and Ubiquitylation Unit, The Sir James Black Centre, College of Life Sciences, University of Dundee, Dow Street, Dundee DD1 5EH, Scotland, U.K."},{"name":"Centre for Gene Regulation and Expression, College of Life Sciences, University of Dundee, Dundee DD1 5EH, Scotland, U.K."}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"288","published-online":{"date-parts":[[2015,3,6]]},"reference":[{"key":"2021112216145608600_B1","doi-asserted-by":"publisher","first-page":"203","DOI":"10.1146\/annurev-biochem-060310-170328","article-title":"The ubiquitin code","volume":"81","author":"Komander","year":"2012","journal-title":"Annu. 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