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dc.contributor.authorBranigan, Emma
dc.contributor.authorPenedo, Carlos
dc.contributor.authorHay, Ronald Thomas
dc.identifier.citationBranigan , E , Penedo , C & Hay , R T 2020 , ' Ubiquitin transfer by a RING E3 ligase occurs from a closed E2~ubiquitin conformation ' , Nature Communications , vol. 11 , 2846 .
dc.identifier.otherPURE: 267667555
dc.identifier.otherPURE UUID: 0a94d2ea-ef18-4146-8cb2-a8e1f36782d6
dc.identifier.otherORCID: /0000-0002-5807-5385/work/75609818
dc.identifier.otherWOS: 000540527100003
dc.identifier.otherScopus: 85086002444
dc.descriptionFunding: Investigator Award from the Wellcome Trust (098391/Z/12/Z) and (217196/Z/19/Z) and a Programme grant from Cancer Research UK (C434/A21747) to R.T.H.; J.C.P. thanks the University of St Andrews for financial support.en
dc.description.abstractBased on extensive structural analysis it was proposed that RING E3 ligases prime the E2~ubiquitin conjugate (E2~Ub) for catalysis by locking it into a closed conformation, where ubiquitin is folded back onto the E2 exposing the restrained thioester bond to attack by substrate nucleophile. However the proposal that the RING dependent closed conformation of E2~Ub represents the active form that mediates ubiquitin transfer has yet to be experimentally tested. To test this hypothesis we use single molecule Förster Resonance Energy Transfer (smFRET) to measure the conformation of a FRET labelled E2~Ub conjugate, which distinguishes between closed and alternative conformations. We describe a real-time FRET assay with a thioester linked E2~Ub conjugate to monitor single ubiquitination events and demonstrate that ubiquitin is transferred to substrate from the closed conformation. These findings are likely to be relevant to all RING E3 catalysed reactions ligating ubiquitin and other ubiquitin-like proteins (Ubls) to substrates.
dc.relation.ispartofNature Communicationsen
dc.rightsCopyright © The Author(s) 2020. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit
dc.subjectQD Chemistryen
dc.subjectQH301 Biologyen
dc.titleUbiquitin transfer by a RING E3 ligase occurs from a closed E2~ubiquitin conformationen
dc.typeJournal articleen
dc.description.versionPublisher PDFen
dc.contributor.institutionUniversity of St Andrews. Centre for Biophotonicsen
dc.contributor.institutionUniversity of St Andrews. Biomedical Sciences Research Complexen
dc.contributor.institutionUniversity of St Andrews. School of Physics and Astronomyen
dc.description.statusPeer revieweden

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