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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Autores: Benítez-Mateos, Ana I., Mehravar, Ehsan, Velasco-Lozano, Susana, Tomovska, Radmila, Salassa, Luca, López-Gallego, Fernando
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2019
País:España
Institución:Universidad de Zaragoza
Repositorio:Zaguán. Repositorio Digital de la Universidad de Zaragoza
OAI Identifier:oai:zaguan.unizar.es:86365
Acceso en línea:http://zaguan.unizar.es/record/86365
Access Level:acceso abierto
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spelling Selective Immobilization of Fluorescent Proteins for the Fabrication of Photoactive MaterialsBenítez-Mateos, Ana I.Mehravar, EhsanVelasco-Lozano, SusanaTomovska, RadmilaSalassa, LucaLópez-Gallego, FernandoThe 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.2019info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://zaguan.unizar.es/record/86365reponame:Zaguán. Repositorio Digital de la Universidad de Zaragozainstname:Universidad de ZaragozaInglésinfo:eu-repo/grantAgreement/ES/MINECO/BIO2014-61838-EXPinfo:eu-repo/grantAgreement/ES/MINECO/BIO2015-69887-Rinfo:eu-repo/grantAgreement/ES/MINECO/CTQ2012-39315info:eu-repo/semantics/openAccessoai:zaguan.unizar.es:863652026-05-29T13:59:51Z
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.
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
Tomovska, Radmila
Salassa, Luca
López-Gallego, Fernando
author Benítez-Mateos, Ana I.
author_facet Benítez-Mateos, Ana I.
Mehravar, Ehsan
Velasco-Lozano, Susana
Tomovska, Radmila
Salassa, Luca
López-Gallego, Fernando
author_role author
author2 Mehravar, Ehsan
Velasco-Lozano, Susana
Tomovska, Radmila
Salassa, Luca
López-Gallego, Fernando
author2_role author
author
author
author
author
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
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://zaguan.unizar.es/record/86365
url http://zaguan.unizar.es/record/86365
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv info:eu-repo/grantAgreement/ES/MINECO/BIO2014-61838-EXP
info:eu-repo/grantAgreement/ES/MINECO/BIO2015-69887-R
info:eu-repo/grantAgreement/ES/MINECO/CTQ2012-39315
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv
publisher.none.fl_str_mv
dc.source.none.fl_str_mv reponame:Zaguán. Repositorio Digital de la Universidad de Zaragoza
instname:Universidad de Zaragoza
instname_str Universidad de Zaragoza
reponame_str Zaguán. Repositorio Digital de la Universidad de Zaragoza
collection Zaguán. Repositorio Digital de la Universidad de Zaragoza
repository.name.fl_str_mv
repository.mail.fl_str_mv
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