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Design and evolution of a synthetic small protein scaffold based on the WW domain

2025-06Artigo científico Acesso aberto
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dc.contributor.authorDias, Ana Margarida Gonçalves Carvalho
dc.contributor.authorTeixeira, Gonçalo Duarte Gomes
dc.contributor.authorBarbosa, Arménio Jorge Moura
dc.contributor.authorGoncalves, Joao
dc.contributor.authorIranzo, Olga
dc.contributor.authorRoque, Ana Cecília Afonso
dc.contributor.institutionUCIBIO - Applied Molecular Biosciences Unit
dc.contributor.institutionDQ - Departamento de Química
dc.contributor.pblWiley-Blackwell
dc.date.accessioned2025-07-29T21:22:31Z
dc.date.available2025-07-29T21:22:31Z
dc.date.issued2025-06
dc.descriptionFunding Information: The authors would like to acknowledge the financial support of FCT—Fundação para a Ciência e a Tecnologia, I.P., in the scope of the project UIDP/04378/2020 and UIDB/04378/2020 of the Research Unit on Applied Molecular Biosciences—UCIBIO and the project LA/P/0140/2020 of the Associate Laboratory Institute for Health and Bioeconomy—i4HB, projects UIDB/04138/2020 e UIDP/04138/2020 of the Research Institute for Medicines (iMed) and Gonçalo Teixeira PhD grant (PD/BD/139800/2018; COVID/BD/152648/2022). This work has received funding from the European Union's Horizon 2020 programme under grant agreement No. 899732 (PURE Project) and from Fundação para a Ciência e Tecnologia (Portugal) and ERDF under the PT2020 Partnership Agreement (LISBOA-01-0145-FEDER-028878) for funding the Sea2See project (PTDC/BII-BIO/28878/2017) and Proteios project (PTDC/CTM-CTM/3389/2021). Publisher Copyright: © 2025 The Author(s). Protein Science published by Wiley Periodicals LLC on behalf of The Protein Society.
dc.description.abstractProtein engineering is a powerful tool in biotechnology and the basis to create unprecedented sequences, structures, and functions. The WW domains are a family of naturally occurring proteins involved in the molecular recognition of proline-rich and phosphorylated peptide sequences with relevance in cellular processes involved in human diseases. Due to their small size, WW domains represent appealing small protein domains for protein engineering and to generate novel functions as binders to non-cognate targets. Here, we designed a synthetic protein scaffold library based on the WW prototype sequence in which the loops were extended and randomized while maintaining structural stability. Using in vitro evolution by phage display against human serum albumin (HSA), we found a lead candidate that was produced by biological and chemical means and further characterized using experimental and computational tools. As a potential application for the lead binder, it was immobilized on a matrix and used to capture the target HSA. Overall, this work shows the versatility of WW domains as peptide scaffolds amenable for in vitro evolution against non-cognate targets.en
dc.description.versionpublishersversion
dc.description.versionpublished
dc.format.extent16
dc.format.extent30445384
dc.identifier.doi10.1002/pro.70164
dc.identifier.issn0961-8368
dc.identifier.otherPURE: 123128713
dc.identifier.otherPURE UUID: 3c67e920-a00b-45a8-8333-d66f52752ecd
dc.identifier.otherScopus: 105005970811
dc.identifier.otherPubMedCentral: PMC12095922
dc.identifier.otherORCID: /0000-0002-9756-4958/work/188867194
dc.identifier.otherORCID: /0000-0001-7416-7215/work/188867970
dc.identifier.urihttp://hdl.handle.net/10362/185705
dc.identifier.urlhttps://www.scopus.com/pages/publications/105005970811
dc.identifier.urlhttps://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=nova_api&SrcAuth=WosAPI&KeyUT=WOS:001492407100001&DestLinkType=FullRecord&DestApp=WOS_CPL
dc.language.isoeng
dc.peerreviewedyes
dc.relationinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F04378%2F2020/PT
dc.relationApplied Molecular Biosciences Unit
dc.relationApplied Molecular Biosciences Unit
dc.relationinfo:eu-repo/grantAgreement/FCT/Concurso para Atribuição do Estatuto e Financiamento de Laboratórios Associados (LA)/LA%2FP%2F0140%2F2020/PT
dc.relationinfo:eu-repo/grantAgreement/FCT/Concurso de avaliação no âmbito do Programa Plurianual de Financiamento de Unidades de I&D (2017%2F2018) - Financiamento Base/UIDB%2F04138%2F2020/PT
dc.relationEngineered proteins for gas sensing
dc.relationinfo:eu-repo/grantAgreement/FCT//PD%2FBD%2F139800%2F2018/PT
dc.relationPrecisely Patterned Nanofibers for High Performance Bioseparations
dc.relationBio-inspired materials for fish spoilage control
dc.relationPROTEIOS - Materiais proteicos para sensorização não invasiva
dc.relationinfo:eu-repo/grantAgreement/FCT/9471 - RIDTI/PTDC%2FBII-BIO%2F28878%2F2017/PT
dc.relationinfo:eu-repo/grantAgreement/FCT/Concurso de Projetos IC&DT em Todos os Domínios Científicos/PTDC%2FCTM-CTM%2F3389%2F2021/PT
dc.subjectAffinity purification
dc.subjectHuman serum albumin
dc.subjectPhage display
dc.subjectProtein engineering
dc.subjectWW domains
dc.subjectBiochemistry
dc.subjectMolecular Biology
dc.subjectSDG 3 - Good Health and Well-being
dc.titleDesign and evolution of a synthetic small protein scaffold based on the WW domainen
dc.typejournal article
degois.publication.firstPage1
degois.publication.issue6
degois.publication.lastPage16
degois.publication.titleProtein Science
degois.publication.volume34
dspace.entity.typePublication
oaire.awardNumberUIDP/04378/2020
oaire.awardNumberUIDB/04378/2020
oaire.awardNumberLA/P/0140/2020
oaire.awardNumberUIDB/04138/2020
oaire.awardNumberUIDP/04138/2020
oaire.awardNumberPD/BD/139800/2018
oaire.awardNumberCOVID/BD/152648/2022
oaire.awardNumber899732
oaire.awardNumberPTDC/BII-BIO/28878/2017
oaire.awardNumberPTDC/CTM-CTM/3389/2021
oaire.awardTitleApplied Molecular Biosciences Unit
oaire.awardTitleApplied Molecular Biosciences Unit
oaire.awardTitleEngineered proteins for gas sensing
oaire.awardTitlePrecisely Patterned Nanofibers for High Performance Bioseparations
oaire.awardTitleBio-inspired materials for fish spoilage control
oaire.awardTitlePROTEIOS - Materiais proteicos para sensorização não invasiva
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F04378%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F04378%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/Concurso para Atribuição do Estatuto e Financiamento de Laboratórios Associados (LA)/LA%2FP%2F0140%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/Concurso de avaliação no âmbito do Programa Plurianual de Financiamento de Unidades de I&D (2017%2F2018) - Financiamento Base/UIDB%2F04138%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/Concurso de avaliação no âmbito do Programa Plurianual de Financiamento de Unidades de I&D (2017%2F2018) - Financiamento Programático/UIDP%2F04138%2F2020/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT//PD%2FBD%2F139800%2F2018/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/OE/COVID%2FBD%2F152648%2F2022/PT
oaire.awardURIinfo:eu-repo/grantAgreement/EC/H2020/899732/EU
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/9471 - RIDTI/PTDC%2FBII-BIO%2F28878%2F2017/PT
oaire.awardURIinfo:eu-repo/grantAgreement/FCT/Concurso de Projetos IC&DT em Todos os Domínios Científicos/PTDC%2FCTM-CTM%2F3389%2F2021/PT
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStream6817 - DCRRNI ID
oaire.fundingStreamConcurso para Atribuição do Estatuto e Financiamento de Laboratórios Associados (LA)
oaire.fundingStreamConcurso de avaliação no âmbito do Programa Plurianual de Financiamento de Unidades de I&D (2017/2018) - Financiamento Base
oaire.fundingStreamConcurso de avaliação no âmbito do Programa Plurianual de Financiamento de Unidades de I&D (2017/2018) - Financiamento Programático
oaire.fundingStreamOE
oaire.fundingStreamH2020
oaire.fundingStream9471 - RIDTI
oaire.fundingStreamConcurso de Projetos IC&DT em Todos os Domínios Científicos
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project.funder.nameEuropean Commission
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