Selective Immobilization of Fluorescent Proteins for the Fabrication of Photoactive Materials

Benítez-Mateos, Ana I. ; Mehravar, Ehsan ; Velasco-Lozano, Susana ; Tomovska, Radmila ; Salassa, Luca ; López-Gallego, Fernando (Universidad de Zaragoza)

Resumen: 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.

Idioma: Inglés

DOI: 10.3390/molecules24152775

Año: 2019

Publicado en: Molecules 24, 15 (2019), 2775 [17 pp.]
ISSN: 1420-3049

Factor impacto JCR: 3.267 (2019)

Categ. JCR: CHEMISTRY, MULTIDISCIPLINARY rank: 70 / 177 = 0.395 (2019) - Q2 - T2
Categ. JCR: BIOCHEMISTRY & MOLECULAR BIOLOGY rank: 141 / 297 = 0.475 (2019) - Q2 - T2

Factor impacto SCIMAGO: 0.698 - Pharmaceutical Science (Q1) - Chemistry (miscellaneous) (Q2) - Drug Discovery (Q2) - Physical and Theoretical Chemistry (Q2) - Organic Chemistry (Q2) - Analytical Chemistry (Q2) - Medicine (miscellaneous) (Q2) - Molecular Medicine (Q3)

Financiación: info:eu-repo/grantAgreement/ES/MINECO/BIO2014-61838-EXP
Financiación: info:eu-repo/grantAgreement/ES/MINECO/BIO2015-69887-R
Financiación: info:eu-repo/grantAgreement/ES/MINECO/CTQ2012-39315

Tipo y forma: Artículo (Versión definitiva)

Área (Departamento): Área Química Orgánica (Dpto. Química Orgánica)