Reversible and irreversible fluorescence activity of the Enhanced Green Fluorescent Protein in pH: Insights for the development of pH-biosensors
Autor(a) principal: | |
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Data de Publicação: | 2020 |
Outros Autores: | , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1016/j.ijbiomac.2020.08.224 http://hdl.handle.net/11449/199340 |
Resumo: | Enhanced Green Fluorescent Protein (EGFP) is a biomolecule with intense and natural fluorescence, with biological and medical applications. Although widely used as a biomarker in research, its application as a biosensor is limited by the lack of in-depth knowledge regarding its structure and behavior in adverse conditions. This study is focused on addressing this need by evaluating EGFP activity and structure at different pH using three-dimensional fluorescence, circular dichroism and small-angle X-ray scattering. The focus was on the reversibility of the process to gain insights for the development of biocompatible pH-biosensors. EGFP was highly stable at alkaline pH and quenched from neutral-to-acidic pH. Above pH 6.0, the fluorescence loss was almost completely reversible on return to neutral pH, but only partially reversible from pH 5.0 to 2.0. This work updates the knowledge regarding EGFP behavior in pH by accounting for the recent data on its structure. Hence, it is evident that EGFP presents the required properties for use as natural, biocompatible and environmentally friendly neutral to acidic pH-biosensors. |
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Reversible and irreversible fluorescence activity of the Enhanced Green Fluorescent Protein in pH: Insights for the development of pH-biosensorsBiosensorCircular dichroism (CD)FluorescenceGreen Fluorescent ProteinIntrinsic fluorescence of proteinsProtein stabilitySmall-angle X-ray scattering (SAXS)Enhanced Green Fluorescent Protein (EGFP) is a biomolecule with intense and natural fluorescence, with biological and medical applications. Although widely used as a biomarker in research, its application as a biosensor is limited by the lack of in-depth knowledge regarding its structure and behavior in adverse conditions. This study is focused on addressing this need by evaluating EGFP activity and structure at different pH using three-dimensional fluorescence, circular dichroism and small-angle X-ray scattering. The focus was on the reversibility of the process to gain insights for the development of biocompatible pH-biosensors. EGFP was highly stable at alkaline pH and quenched from neutral-to-acidic pH. Above pH 6.0, the fluorescence loss was almost completely reversible on return to neutral pH, but only partially reversible from pH 5.0 to 2.0. This work updates the knowledge regarding EGFP behavior in pH by accounting for the recent data on its structure. Hence, it is evident that EGFP presents the required properties for use as natural, biocompatible and environmentally friendly neutral to acidic pH-biosensors.Australian Nuclear Science and Technology OrganisationCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Department of Engineering of Bioprocesses and Biotechnology School of Pharmaceutical Sciences São Paulo State University (UNESP), Rodovia Araraquara-Jaú/Km 01School of Science College of Science Engineering and Health RMIT University, 124 La Trobe StreetAustralian Synchrotron, 800 Blackburn RoadUniv Coimbra CIEPQPF Department of Chemical Engineering, Rua Sílvio Lima, Pólo II – Pinhal de MarrocosDepartment of Engineering of Bioprocesses and Biotechnology School of Pharmaceutical Sciences São Paulo State University (UNESP), Rodovia Araraquara-Jaú/Km 01FAPESP: 18/06576-5FAPESP: 2014/16424-7FAPESP: 2014/19793-3FAPESP: 2016/07529-5FAPESP: 2018/01858-2FAPESP: 2018/20833-0FAPESP: 2018/50009-8Universidade Estadual Paulista (Unesp)RMIT UniversityAustralian SynchrotronCIEPQPFdos Santos, Nathalia Vieira [UNESP]Saponi, Carolina Falaschi [UNESP]Ryan, Timothy M.Primo, Fernando L. [UNESP]Greaves, Tamar L.Pereira, Jorge F.B. [UNESP]2020-12-12T01:37:10Z2020-12-12T01:37:10Z2020-12-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article3474-3484http://dx.doi.org/10.1016/j.ijbiomac.2020.08.224International Journal of Biological Macromolecules, v. 164, p. 3474-3484.1879-00030141-8130http://hdl.handle.net/11449/19934010.1016/j.ijbiomac.2020.08.2242-s2.0-85090277964Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengInternational Journal of Biological Macromoleculesinfo:eu-repo/semantics/openAccess2021-10-23T07:21:25Zoai:repositorio.unesp.br:11449/199340Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462021-10-23T07:21:25Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Reversible and irreversible fluorescence activity of the Enhanced Green Fluorescent Protein in pH: Insights for the development of pH-biosensors |
title |
Reversible and irreversible fluorescence activity of the Enhanced Green Fluorescent Protein in pH: Insights for the development of pH-biosensors |
spellingShingle |
Reversible and irreversible fluorescence activity of the Enhanced Green Fluorescent Protein in pH: Insights for the development of pH-biosensors dos Santos, Nathalia Vieira [UNESP] Biosensor Circular dichroism (CD) Fluorescence Green Fluorescent Protein Intrinsic fluorescence of proteins Protein stability Small-angle X-ray scattering (SAXS) |
title_short |
Reversible and irreversible fluorescence activity of the Enhanced Green Fluorescent Protein in pH: Insights for the development of pH-biosensors |
title_full |
Reversible and irreversible fluorescence activity of the Enhanced Green Fluorescent Protein in pH: Insights for the development of pH-biosensors |
title_fullStr |
Reversible and irreversible fluorescence activity of the Enhanced Green Fluorescent Protein in pH: Insights for the development of pH-biosensors |
title_full_unstemmed |
Reversible and irreversible fluorescence activity of the Enhanced Green Fluorescent Protein in pH: Insights for the development of pH-biosensors |
title_sort |
Reversible and irreversible fluorescence activity of the Enhanced Green Fluorescent Protein in pH: Insights for the development of pH-biosensors |
author |
dos Santos, Nathalia Vieira [UNESP] |
author_facet |
dos Santos, Nathalia Vieira [UNESP] Saponi, Carolina Falaschi [UNESP] Ryan, Timothy M. Primo, Fernando L. [UNESP] Greaves, Tamar L. Pereira, Jorge F.B. [UNESP] |
author_role |
author |
author2 |
Saponi, Carolina Falaschi [UNESP] Ryan, Timothy M. Primo, Fernando L. [UNESP] Greaves, Tamar L. Pereira, Jorge F.B. [UNESP] |
author2_role |
author author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) RMIT University Australian Synchrotron CIEPQPF |
dc.contributor.author.fl_str_mv |
dos Santos, Nathalia Vieira [UNESP] Saponi, Carolina Falaschi [UNESP] Ryan, Timothy M. Primo, Fernando L. [UNESP] Greaves, Tamar L. Pereira, Jorge F.B. [UNESP] |
dc.subject.por.fl_str_mv |
Biosensor Circular dichroism (CD) Fluorescence Green Fluorescent Protein Intrinsic fluorescence of proteins Protein stability Small-angle X-ray scattering (SAXS) |
topic |
Biosensor Circular dichroism (CD) Fluorescence Green Fluorescent Protein Intrinsic fluorescence of proteins Protein stability Small-angle X-ray scattering (SAXS) |
description |
Enhanced Green Fluorescent Protein (EGFP) is a biomolecule with intense and natural fluorescence, with biological and medical applications. Although widely used as a biomarker in research, its application as a biosensor is limited by the lack of in-depth knowledge regarding its structure and behavior in adverse conditions. This study is focused on addressing this need by evaluating EGFP activity and structure at different pH using three-dimensional fluorescence, circular dichroism and small-angle X-ray scattering. The focus was on the reversibility of the process to gain insights for the development of biocompatible pH-biosensors. EGFP was highly stable at alkaline pH and quenched from neutral-to-acidic pH. Above pH 6.0, the fluorescence loss was almost completely reversible on return to neutral pH, but only partially reversible from pH 5.0 to 2.0. This work updates the knowledge regarding EGFP behavior in pH by accounting for the recent data on its structure. Hence, it is evident that EGFP presents the required properties for use as natural, biocompatible and environmentally friendly neutral to acidic pH-biosensors. |
publishDate |
2020 |
dc.date.none.fl_str_mv |
2020-12-12T01:37:10Z 2020-12-12T01:37:10Z 2020-12-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1016/j.ijbiomac.2020.08.224 International Journal of Biological Macromolecules, v. 164, p. 3474-3484. 1879-0003 0141-8130 http://hdl.handle.net/11449/199340 10.1016/j.ijbiomac.2020.08.224 2-s2.0-85090277964 |
url |
http://dx.doi.org/10.1016/j.ijbiomac.2020.08.224 http://hdl.handle.net/11449/199340 |
identifier_str_mv |
International Journal of Biological Macromolecules, v. 164, p. 3474-3484. 1879-0003 0141-8130 10.1016/j.ijbiomac.2020.08.224 2-s2.0-85090277964 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
International Journal of Biological Macromolecules |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
3474-3484 |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
_version_ |
1803046440409235456 |