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dc.contributor.authorYadahalli, Shilpaen
dc.contributor.authorNeira, José Len
dc.contributor.authorJohnson, Christopher Men
dc.contributor.authorTan, Yaw Singen
dc.contributor.authorRowling, Pamelaen
dc.contributor.authorChattopadhyay, Anasuyaen
dc.contributor.authorVerma, Chandra Sen
dc.contributor.authorItzhaki, Lauraen
dc.date.accessioned2019-01-30T00:30:15Z
dc.date.available2019-01-30T00:30:15Z
dc.date.issued2019-01-24en
dc.identifier.issn2045-2322
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/288489
dc.description.abstractp53 is frequently mutated in human cancers. Its levels are tightly regulated by the E3 ubiquitin ligase MDM2. The complex between MDM2 and p53 is largely formed by the interaction between the N-terminal domain of MDM2 and the N-terminal transactivation (TA) domain of p53 (residues 15-29). We investigated the kinetic and thermodynamic basis of the MDM2/p53 interaction by using wild-type and mutant variants of the TA domain. We focus on the effects of phosphorylation at positions Thr18 and Ser20 including their substitution with phosphomimetics. Conformational propensities of the isolated peptides were investigated using in silico methods and experimentally by circular dichroism and 1H-NMR in aqueous solution. Both experimental and computational analyses indicate that the p53 peptides are mainly disordered in aqueous solution, with evidence of nascent helix around the Ser20-Leu25 region. Both phosphorylation and the phosphomimetics at Thr18 resulted in a decrease in the binding affinity by ten- to twenty-fold when compared to the wild-type. Phosphorylation and phosphomimetics at Ser20 resulted in a smaller decrease in the affinity. Mutation of residues Lys24 and Leu25 also disrupted the interaction. Our results may be useful for further development of peptide-based drugs targeting the MDM2/p53 interaction.
dc.description.sponsorshipGeneralitat Valenciana BEST short-stay fellowship in the Department of Pharmacology, Cambridge, UK. Medical Research Foundation
dc.format.mediumElectronicen
dc.languageengen
dc.publisherNature Publishing Group
dc.rightsAttribution 4.0 International
dc.rights.urihttps://creativecommons.org/licenses/by/4.0/
dc.subjectHumansen
dc.subjectThreonineen
dc.subjectSerineen
dc.subjectPeptide Fragmentsen
dc.subjectSpectrometry, Fluorescenceen
dc.subjectCircular Dichroismen
dc.subjectMagnetic Resonance Spectroscopyen
dc.subjectBinding Sitesen
dc.subjectProtein Conformationen
dc.subjectPhosphorylationen
dc.subjectKineticsen
dc.subjectMutationen
dc.subjectThermodynamicsen
dc.subjectTumor Suppressor Protein p53en
dc.subjectProto-Oncogene Proteins c-mdm2en
dc.subjectProtein Interaction Domains and Motifsen
dc.subjectMolecular Dynamics Simulationen
dc.titleKinetic and thermodynamic effects of phosphorylation on p53 binding to MDM2.en
dc.typeArticle
prism.issueIdentifier1en
prism.publicationDate2019en
prism.publicationNameScientific reportsen
prism.startingPage693
prism.volume9en
dc.identifier.doi10.17863/CAM.35776
dcterms.dateAccepted2018-11-15en
rioxxterms.versionofrecord10.1038/s41598-018-36589-5en
rioxxterms.versionVoR
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2019-01-24en
dc.contributor.orcidTan, Yaw Sing [0000-0002-2522-9421]
dc.contributor.orcidItzhaki, Laura [0000-0001-6504-2576]
dc.identifier.eissn2045-2322
rioxxterms.typeJournal Article/Reviewen


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Attribution 4.0 International
Except where otherwise noted, this item's licence is described as Attribution 4.0 International