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dc.contributor.authorFerreira, Ana V.por
dc.contributor.authorAntunes, Egiptopor
dc.contributor.authorRibeiro, Arturpor
dc.contributor.authorMatamá, Teresapor
dc.contributor.authorAzoia, Nuno G.por
dc.contributor.authorCunha, J.por
dc.contributor.authorCavaco-Paulo, Arturpor
dc.identifier.citationFerreira, 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), 2019por
dc.description.abstractRecent 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.por
dc.description.sponsorshipWe 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.por
dc.publisherElsevier BVpor
dc.subjectHuman neutrophil elastase (HNE)por
dc.subjectSplit GFPpor
dc.subjectChromogenic GFP-like proteinspor
dc.subjectUltramarine (UM)por
dc.subjectMolecular dynamics simulationspor
dc.titleDesign of a chromogenic substrate for elastase based on split GFP system proof of concept for colour switch sensorspor
sdum.journalBiotechnology Reportspor
Appears in Collections:CEB - Publicações em Revistas/Séries Internacionais / Publications in International Journals/Series

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