Design of a chromogenic substrate for elastase based on split GFP system proof of concept for colour switch sensors
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Outros Autores: | , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/1822/60429 |
Resumo: | Recent studies have demonstrated that human neutrophil elastase (HNE) can be used as marker for inflammation/infection of chronic wounds since it was found to be present in high concentration in exudate collected from chronic wounds. Biosensors used in wound care benefit from a chromogenic signalling due to the readiness of signal interpretation, but the most common use faint yellow chromogenic molecules such as p-nitroaniline (pNa). In addition, if to be converted into smart dressings, the colour of the detection system should not be masked by the exudates colour. In this work, we designed a chromogenic substrate for HNE aiming to be incorporated in a smart dressing as a colour switch sensor. The substrate was developed using the GFP-like chromoprotein ultramarine (UM), following the split GFP technology. The cleavage sequence for HNE (Ala-Ala-Pro-Val) was embedded into the sensing moiety of the substrate corresponding to the 11th -sheet. In the presence of HNE, the 11th -sheet is able to interact to the signalling moiety composed of the 1-10 incomplete barrel, allowing the re-establishment of the chromophore environment and, hence, the colour production. Structural homology and molecular dynamics simulations were conducted to aid on the disclosure of the structural changes that are the base of the mechanism of action of this HNE switch substrate. Our findings explore the possible application of GFP-like chromogenic sensors in point-of-care devices for the evaluation of the wounds status, representing a major step in the medical field. |
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Design of a chromogenic substrate for elastase based on split GFP system proof of concept for colour switch sensorsHuman neutrophil elastase (HNE)Split GFPChromogenic GFP-like proteinsUltramarine (UM)ProteasesMolecular dynamics simulationsRecent studies have demonstrated that human neutrophil elastase (HNE) can be used as marker for inflammation/infection of chronic wounds since it was found to be present in high concentration in exudate collected from chronic wounds. Biosensors used in wound care benefit from a chromogenic signalling due to the readiness of signal interpretation, but the most common use faint yellow chromogenic molecules such as p-nitroaniline (pNa). In addition, if to be converted into smart dressings, the colour of the detection system should not be masked by the exudates colour. In this work, we designed a chromogenic substrate for HNE aiming to be incorporated in a smart dressing as a colour switch sensor. The substrate was developed using the GFP-like chromoprotein ultramarine (UM), following the split GFP technology. The cleavage sequence for HNE (Ala-Ala-Pro-Val) was embedded into the sensing moiety of the substrate corresponding to the 11th -sheet. In the presence of HNE, the 11th -sheet is able to interact to the signalling moiety composed of the 1-10 incomplete barrel, allowing the re-establishment of the chromophore environment and, hence, the colour production. Structural homology and molecular dynamics simulations were conducted to aid on the disclosure of the structural changes that are the base of the mechanism of action of this HNE switch substrate. Our findings explore the possible application of GFP-like chromogenic sensors in point-of-care devices for the evaluation of the wounds status, representing a major step in the medical field.We greatly acknowledge the European project InFact – Functional materials for fast diagnosis of wound infection (FP7-NMP-2013-SME-7 - Grant agreement no. 604278) for funding the work and also the grant for Joana Cunha and José Egipto Antunes. Ana Ferreira, Artur Ribeiro and Teresa Matamá would also like to acknowledge the Portuguese Foundation for Science and Technology (FCT) for funding their scholarships with the references SFRH/BD/113247/2015, SFRH/BPD/98388/2013 and SFRH/102153/2014, respectively. The work done at Centre of Biological Engineering (CEB) was supported by FCT under the scope of the strategic funding of UID/BIO/04469/2013 unit, COMPETE 2020 (POCI-01-0145-FEDER-006684) and BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte.info:eu-repo/semantics/publishedVersionElsevier BVUniversidade do MinhoFerreira, Ana V.Antunes, EgiptoRibeiro, ArturMatamá, TeresaAzoia, Nuno G.Cunha, J.Cavaco-Paulo, Artur2019-062019-06-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/1822/60429engFerreira, Ana V.; Antunes, Egipto; Ribeiro, Artur; Matamá, Teresa; Azoia, Nuno G.; Cunha, J.; Cavaco-Paulo, Artur, Design of a chromogenic substrate for elastase based on split GFP system proof of concept for colour switch sensors. Biotechnology Reports, 22(e00324), 20192215-017X2215-017X10.1016/j.btre.2019.e00324https://www.journals.elsevier.com/biotechnology-reportsinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T12:45:49Zoai:repositorium.sdum.uminho.pt:1822/60429Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:43:43.939398Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Design of a chromogenic substrate for elastase based on split GFP system proof of concept for colour switch sensors |
title |
Design of a chromogenic substrate for elastase based on split GFP system proof of concept for colour switch sensors |
spellingShingle |
Design of a chromogenic substrate for elastase based on split GFP system proof of concept for colour switch sensors Ferreira, Ana V. Human neutrophil elastase (HNE) Split GFP Chromogenic GFP-like proteins Ultramarine (UM) Proteases Molecular dynamics simulations |
title_short |
Design of a chromogenic substrate for elastase based on split GFP system proof of concept for colour switch sensors |
title_full |
Design of a chromogenic substrate for elastase based on split GFP system proof of concept for colour switch sensors |
title_fullStr |
Design of a chromogenic substrate for elastase based on split GFP system proof of concept for colour switch sensors |
title_full_unstemmed |
Design of a chromogenic substrate for elastase based on split GFP system proof of concept for colour switch sensors |
title_sort |
Design of a chromogenic substrate for elastase based on split GFP system proof of concept for colour switch sensors |
author |
Ferreira, Ana V. |
author_facet |
Ferreira, Ana V. Antunes, Egipto Ribeiro, Artur Matamá, Teresa Azoia, Nuno G. Cunha, J. Cavaco-Paulo, Artur |
author_role |
author |
author2 |
Antunes, Egipto Ribeiro, Artur Matamá, Teresa Azoia, Nuno G. Cunha, J. Cavaco-Paulo, Artur |
author2_role |
author author author author author author |
dc.contributor.none.fl_str_mv |
Universidade do Minho |
dc.contributor.author.fl_str_mv |
Ferreira, Ana V. Antunes, Egipto Ribeiro, Artur Matamá, Teresa Azoia, Nuno G. Cunha, J. Cavaco-Paulo, Artur |
dc.subject.por.fl_str_mv |
Human neutrophil elastase (HNE) Split GFP Chromogenic GFP-like proteins Ultramarine (UM) Proteases Molecular dynamics simulations |
topic |
Human neutrophil elastase (HNE) Split GFP Chromogenic GFP-like proteins Ultramarine (UM) Proteases Molecular dynamics simulations |
description |
Recent studies have demonstrated that human neutrophil elastase (HNE) can be used as marker for inflammation/infection of chronic wounds since it was found to be present in high concentration in exudate collected from chronic wounds. Biosensors used in wound care benefit from a chromogenic signalling due to the readiness of signal interpretation, but the most common use faint yellow chromogenic molecules such as p-nitroaniline (pNa). In addition, if to be converted into smart dressings, the colour of the detection system should not be masked by the exudates colour. In this work, we designed a chromogenic substrate for HNE aiming to be incorporated in a smart dressing as a colour switch sensor. The substrate was developed using the GFP-like chromoprotein ultramarine (UM), following the split GFP technology. The cleavage sequence for HNE (Ala-Ala-Pro-Val) was embedded into the sensing moiety of the substrate corresponding to the 11th -sheet. In the presence of HNE, the 11th -sheet is able to interact to the signalling moiety composed of the 1-10 incomplete barrel, allowing the re-establishment of the chromophore environment and, hence, the colour production. Structural homology and molecular dynamics simulations were conducted to aid on the disclosure of the structural changes that are the base of the mechanism of action of this HNE switch substrate. Our findings explore the possible application of GFP-like chromogenic sensors in point-of-care devices for the evaluation of the wounds status, representing a major step in the medical field. |
publishDate |
2019 |
dc.date.none.fl_str_mv |
2019-06 2019-06-01T00:00:00Z |
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://hdl.handle.net/1822/60429 |
url |
http://hdl.handle.net/1822/60429 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Ferreira, Ana V.; Antunes, Egipto; Ribeiro, Artur; Matamá, Teresa; Azoia, Nuno G.; Cunha, J.; Cavaco-Paulo, Artur, Design of a chromogenic substrate for elastase based on split GFP system proof of concept for colour switch sensors. Biotechnology Reports, 22(e00324), 2019 2215-017X 2215-017X 10.1016/j.btre.2019.e00324 https://www.journals.elsevier.com/biotechnology-reports |
dc.rights.driver.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 |
Elsevier BV |
publisher.none.fl_str_mv |
Elsevier BV |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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