Selective immobilization of fluorescent proteins for the fabrication of photoactive materials

The immobilization of fluorescent proteins is a key technology enabling to fabricate a new generation of photoactive materials with potential technological applications. Herein we have exploited superfolder green (sGFP) and red (RFP) fluorescent proteins expressed with different polypeptide tags. We...

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Detalles Bibliográficos
Autores: Benítez-Mateos, Ana I., Mehravar, Ehsan, Velasco-Lozano, Susana, Salassa, Luca, López-Gallego, Fernando
Tipo de recurso: artículo
Fecha de publicación:2019
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/208569
Acceso en línea:http://hdl.handle.net/10261/208569
Access Level:acceso abierto
Palabra clave:protein immobilization
polypeptide-tags
fluorescent proteins
upconverting
nanomaterials
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spelling Selective immobilization of fluorescent proteins for the fabrication of photoactive materialsBenítez-Mateos, Ana I.Mehravar, EhsanVelasco-Lozano, SusanaSalassa, LucaLópez-Gallego, Fernandoprotein immobilizationpolypeptide-tagsfluorescent proteinsupconvertingnanomaterialsThe immobilization of fluorescent proteins is a key technology enabling to fabricate a new generation of photoactive materials with potential technological applications. Herein we have exploited superfolder green (sGFP) and red (RFP) fluorescent proteins expressed with different polypeptide tags. We fused these fluorescent proteins to His-tags to immobilize them on graphene 3D hydrogels, and Cys-tags to immobilize them on porous microparticles activated with either epoxy or disulfide groups and with Lys-tags to immobilize them on upconverting nanoparticles functionalized with carboxylic groups. Genetically programming sGFP and RFP with Cys-tag and His-tag, respectively, allowed tuning the protein spatial organization either across the porous structure of two microbeads with different functional groups (agarose-based materials activated with metal chelates and epoxy-methacrylate materials) or across the surface of a single microbead functionalized with both metal-chelates and disulfide groups. By using different polypeptide tags, we can control the attachment chemistry but also the localization of the fluorescent proteins across the material surfaces. The resulting photoactive material formed by His-RFP immobilized on graphene hydrogels has been tested as pH indicator to measure pH changes in the alkaline region, although the immobilized fluorescent protein exhibited a narrower dynamic range to measure pH than the soluble fluorescent protein. Likewise, the immobilization of Lys-sGFP on alginate-coated upconverting nanoparticles enabled the infrared excitation of the fluorescent protein to be used as a green light emitter. These novel photoactive biomaterials open new avenues for innovative technological developments towards the fabrication of biosensors and photonic devices.A.I.B.-M. and F.L.-G. are grateful to MINECO (BIO2015-69887-R and BIO2014-61838-EXP) for funding them. We also thank ARAID foundation for funding F.L.-G., and S.V.-L. thanks the Mexican Council of Science and Technology (CONACyT) for the postdoctoral fellowship she received. L.S. thanks MINECO for grant CTQ2012-39315.Molecular Diversity Preservation InternationalMinisterio de Economía y Competitividad (España)ARAID FoundationConsejo Nacional de Ciencia y Tecnología (México)Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]2020202020192020info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501http://hdl.handle.net/10261/208569reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2015-69887-Rinfo:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2014-61838-EXPhttp://doi.org/10.3390/molecules24152775Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/2085692026-05-22T06:33:51Z
dc.title.none.fl_str_mv Selective immobilization of fluorescent proteins for the fabrication of photoactive materials
title Selective immobilization of fluorescent proteins for the fabrication of photoactive materials
spellingShingle Selective immobilization of fluorescent proteins for the fabrication of photoactive materials
Benítez-Mateos, Ana I.
protein immobilization
polypeptide-tags
fluorescent proteins
upconverting
nanomaterials
title_short Selective immobilization of fluorescent proteins for the fabrication of photoactive materials
title_full Selective immobilization of fluorescent proteins for the fabrication of photoactive materials
title_fullStr Selective immobilization of fluorescent proteins for the fabrication of photoactive materials
title_full_unstemmed Selective immobilization of fluorescent proteins for the fabrication of photoactive materials
title_sort Selective immobilization of fluorescent proteins for the fabrication of photoactive materials
dc.creator.none.fl_str_mv Benítez-Mateos, Ana I.
Mehravar, Ehsan
Velasco-Lozano, Susana
Salassa, Luca
López-Gallego, Fernando
author Benítez-Mateos, Ana I.
author_facet Benítez-Mateos, Ana I.
Mehravar, Ehsan
Velasco-Lozano, Susana
Salassa, Luca
López-Gallego, Fernando
author_role author
author2 Mehravar, Ehsan
Velasco-Lozano, Susana
Salassa, Luca
López-Gallego, Fernando
author2_role author
author
author
author
dc.contributor.none.fl_str_mv Ministerio de Economía y Competitividad (España)
ARAID Foundation
Consejo Nacional de Ciencia y Tecnología (México)
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv protein immobilization
polypeptide-tags
fluorescent proteins
upconverting
nanomaterials
topic protein immobilization
polypeptide-tags
fluorescent proteins
upconverting
nanomaterials
description The immobilization of fluorescent proteins is a key technology enabling to fabricate a new generation of photoactive materials with potential technological applications. Herein we have exploited superfolder green (sGFP) and red (RFP) fluorescent proteins expressed with different polypeptide tags. We fused these fluorescent proteins to His-tags to immobilize them on graphene 3D hydrogels, and Cys-tags to immobilize them on porous microparticles activated with either epoxy or disulfide groups and with Lys-tags to immobilize them on upconverting nanoparticles functionalized with carboxylic groups. Genetically programming sGFP and RFP with Cys-tag and His-tag, respectively, allowed tuning the protein spatial organization either across the porous structure of two microbeads with different functional groups (agarose-based materials activated with metal chelates and epoxy-methacrylate materials) or across the surface of a single microbead functionalized with both metal-chelates and disulfide groups. By using different polypeptide tags, we can control the attachment chemistry but also the localization of the fluorescent proteins across the material surfaces. The resulting photoactive material formed by His-RFP immobilized on graphene hydrogels has been tested as pH indicator to measure pH changes in the alkaline region, although the immobilized fluorescent protein exhibited a narrower dynamic range to measure pH than the soluble fluorescent protein. Likewise, the immobilization of Lys-sGFP on alginate-coated upconverting nanoparticles enabled the infrared excitation of the fluorescent protein to be used as a green light emitter. These novel photoactive biomaterials open new avenues for innovative technological developments towards the fabrication of biosensors and photonic devices.
publishDate 2019
dc.date.none.fl_str_mv 2019
2020
2020
2020
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
format article
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/208569
url http://hdl.handle.net/10261/208569
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv #PLACEHOLDER_PARENT_METADATA_VALUE#
#PLACEHOLDER_PARENT_METADATA_VALUE#
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2015-69887-R
info:eu-repo/grantAgreement/MINECO/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/BIO2014-61838-EXP
http://doi.org/10.3390/molecules24152775
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Molecular Diversity Preservation International
publisher.none.fl_str_mv Molecular Diversity Preservation International
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
repository.mail.fl_str_mv
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score 15.811543

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