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oai:uvadoc.uva.es:10324/229072021-06-24T07:27:12Zcom_10324_22821com_10324_954com_10324_894col_10324_22822
UVaDOC author Ibáñez Fonseca, Arturo author Alonso Rodrigo, Matilde author Arias Vallejo, Francisco Javier author Rodríguez Cabello, José Carlos 2017-03-31T06:46:08Z 2017-03-31T06:46:08Z 2017 Bioconjugate Chem., 2017, 28 (3), pp 828–835 http://uvadoc.uva.es/handle/10324/22907 10.1021/acs.bioconjchem.6b00738 In the last decades, recombinant structural proteins have become very promising in addressing different issues such as the lack of traceability of biomedical devices or the design of more sensitive biosensors. Among them, we find elastin-like recombinamers (ELRs), which can be designed to self-assemble into diverse structures, such as hydrogels. Furthermore, they might be combined with other protein polymers, such as silk, to give silk-elastin-like recombinamers (SELRs), holding the properties of both proteins. In this work, due to their recombinant nature, we have fused two different fluorescent proteins (FPs), i.e., the green Aequorea coerulescens enhanced green fluorescent protein and the near-infrared eqFP650, to a SELR able to form irreversible hydrogels through physical cross-linking. These recombinamers showed an emission of fluorescence similar to the single FPs, and they were capable of forming hydrogels with different stiffness (G′ = 60–4000 Pa) by varying the concentration of the SELR-FPs. Moreover, the absorption spectrum of SELR-eqFP650 showed a peak greatly overlapping the emission spectrum of the SELR-Aequorea coerulescens enhanced green fluorescent protein. Hence, this enables Förster resonance energy transfer (FRET) upon the interaction between two SELR molecules, each one containing a different FP, due to the stacking of silk domains at any temperature and to the aggregation of elastin-like blocks above the transition temperature. This effect was studied by different methods, and a FRET efficiency of 0.06–0.2 was observed, depending on the technique used for its calculation. Therefore, innovative biological applications arise from the combination of SELRs with FPs, such as enhancing the traceability of hydrogels based on SELRs intended for tissue engineering, the development of biosensors, and the prediction of FRET efficiencies of novel FRET pairs. eng FRET-Paired Hydrogel Forming Silk-Elastin-Like Recombinamers by Recombinant Conjugation of Fluorescent Proteins info:eu-repo/semantics/article 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URL https://uvadoc.uva.es/bitstream/10324/22907/4/FRET-paired%20hydrogel%20forming%20silk-elastin-like%20recombinamers.pdf File MD5 9b6a80b4fe3050d5560f390f09dec661 762505 application/pdf FRET-paired hydrogel forming silk-elastin-like recombinamers.pdf URL https://uvadoc.uva.es/bitstream/10324/22907/5/SI_FRET-paired%20hydrogel%20forming%20silk-elastin-like%20recombinamers.pdf File MD5 e8f11233bf19cd6ca3f374fb1e955313 565117 application/pdf SI_FRET-paired hydrogel forming silk-elastin-like recombinamers.pdf