Selective detection of active extracellular granzyme A by using a novel fluorescent immunoprobe with application to inflammatory diseases

8 figures.-- Supplementary information available.

Detalles Bibliográficos
Autores: Senán Salinas, Ana, Comas, Laura, Esteban, Patricia, Garzón, Marcela, Cheng, Zhiming, Santiago, Llipsy, Domingo, María Pilar, Ramírez-Labrada, Ariel, Paño, José Ramón, Vendrell, Marc, Pardo, Julián, Arias, Maykel, Gálvez Buerba, Eva Mª
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2024
País:España
Institución:Consejo Superior de Investigaciones Científicas (CSIC)
Repositorio:DIGITAL.CSIC. Repositorio Institucional del CSIC
OAI Identifier:oai:digital.csic.es:10261/354919
Acceso en línea:http://hdl.handle.net/10261/354919
Access Level:acceso abierto
Palabra clave:GzmA activity
Probe
Fluorescence
Inflammatory disease
http://metadata.un.org/sdg/3
Ensure healthy lives and promote well-being for all at all ages
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oai_identifier_str oai:digital.csic.es:10261/354919
network_acronym_str ES
network_name_str España
repository_id_str
dc.title.none.fl_str_mv Selective detection of active extracellular granzyme A by using a novel fluorescent immunoprobe with application to inflammatory diseases
title Selective detection of active extracellular granzyme A by using a novel fluorescent immunoprobe with application to inflammatory diseases
spellingShingle Selective detection of active extracellular granzyme A by using a novel fluorescent immunoprobe with application to inflammatory diseases
Senán Salinas, Ana
GzmA activity
Probe
Fluorescence
Inflammatory disease
http://metadata.un.org/sdg/3
Ensure healthy lives and promote well-being for all at all ages
title_short Selective detection of active extracellular granzyme A by using a novel fluorescent immunoprobe with application to inflammatory diseases
title_full Selective detection of active extracellular granzyme A by using a novel fluorescent immunoprobe with application to inflammatory diseases
title_fullStr Selective detection of active extracellular granzyme A by using a novel fluorescent immunoprobe with application to inflammatory diseases
title_full_unstemmed Selective detection of active extracellular granzyme A by using a novel fluorescent immunoprobe with application to inflammatory diseases
title_sort Selective detection of active extracellular granzyme A by using a novel fluorescent immunoprobe with application to inflammatory diseases
dc.creator.none.fl_str_mv Senán Salinas, Ana
Comas, Laura
Esteban, Patricia
Garzón, Marcela
Cheng, Zhiming
Santiago, Llipsy
Domingo, María Pilar
Ramírez-Labrada, Ariel
Paño, José Ramón
Vendrell, Marc
Pardo, Julián
Arias, Maykel
Gálvez Buerba, Eva Mª
author Senán Salinas, Ana
author_facet Senán Salinas, Ana
Comas, Laura
Esteban, Patricia
Garzón, Marcela
Cheng, Zhiming
Santiago, Llipsy
Domingo, María Pilar
Ramírez-Labrada, Ariel
Paño, José Ramón
Vendrell, Marc
Pardo, Julián
Arias, Maykel
Gálvez Buerba, Eva Mª
author_role author
author2 Comas, Laura
Esteban, Patricia
Garzón, Marcela
Cheng, Zhiming
Santiago, Llipsy
Domingo, María Pilar
Ramírez-Labrada, Ariel
Paño, José Ramón
Vendrell, Marc
Pardo, Julián
Arias, Maykel
Gálvez Buerba, Eva Mª
author2_role author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Instituto de Salud Carlos III
Ministerio de Ciencia, Innovación y Universidades (España)
European Commission
Gobierno de Aragón
Agencia Estatal de Investigación (España)
European Research Council
Ministerio de Economía y Competitividad (España)
Senán Salinas, Ana [0009-0009-5597-851X]
Comas, Laura [0000-0002-3843-1231]
Esteban, Patricia [0000-0003-4123-3524]
Garzón, Marcela [0000-0001-6778-0636]
Santiago, Llipsy [0000-0002-1861-5981]
Domingo, María Pilar [0000-0002-6829-8769]
Ramírez-Labrada, Ariel [0000-0002-3888-7036]
Paño, José Ramón [0000-0002-9600-8116]
Vendrell, Marc [0000-0002-5392-9740]
Pardo, Julián [0000-0003-0154-0730]
Arias, Maykel [0000-0002-9730-2210]
Gálvez Buerba, Eva Mª [0000-0001-6928-5516]
Gálvez Buerba, Eva Mª [eva@icb.csic.es]
Arias, Maykel [maykelariascabrero@gmail.com]
Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]
dc.subject.none.fl_str_mv GzmA activity
Probe
Fluorescence
Inflammatory disease
http://metadata.un.org/sdg/3
Ensure healthy lives and promote well-being for all at all ages
topic GzmA activity
Probe
Fluorescence
Inflammatory disease
http://metadata.un.org/sdg/3
Ensure healthy lives and promote well-being for all at all ages
description 8 figures.-- Supplementary information available.
publishDate 2024
dc.date.none.fl_str_mv 2024
2024
2024
dc.type.none.fl_str_mv info:eu-repo/semantics/article
http://purl.org/coar/resource_type/c_6501
Publisher's version
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/10261/354919
url http://hdl.handle.net/10261/354919
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
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info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2020-113963RB-I00
info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/SAF2017-83120-C2-1-R
info:eu-repo/grantAgreement/EC/H2020/771443
info:eu-repo/grantAgreement/EC/H2020/859908
Senán Salinas, Ana; Comas, Laura; Esteban, Patricia; Garzón, Marcela; Cheng, Zhiming; Santiago, Llipsy; Domingo, María Pilar; Ramírez-Labrada, Ariel; Paño, José Ramón; Vendrell, Marc; Pardo, Julián; Arias, Maykel; Gálvez Buerba, Eva Mª; 2024; Supporting information for Selective detection of active extracellular granzyme A by using a novel fluorescent immunoprobe with application to inflammatory diseases [Dataset]; ACS Publications; http://dx.doi.org/10.1021/acsptsci.4c00065
https://www.wikidata.org/wiki/Q18026722
http://dx.doi.org/10.1021/acsptsci.4c00065
Sí
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
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dc.publisher.none.fl_str_mv ACS Publications
publisher.none.fl_str_mv ACS Publications
dc.source.none.fl_str_mv reponame:DIGITAL.CSIC. Repositorio Institucional del CSIC
instname:Consejo Superior de Investigaciones Científicas (CSIC)
instname_str Consejo Superior de Investigaciones Científicas (CSIC)
reponame_str DIGITAL.CSIC. Repositorio Institucional del CSIC
collection DIGITAL.CSIC. Repositorio Institucional del CSIC
repository.name.fl_str_mv
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spelling Selective detection of active extracellular granzyme A by using a novel fluorescent immunoprobe with application to inflammatory diseasesSenán Salinas, AnaComas, LauraEsteban, PatriciaGarzón, MarcelaCheng, ZhimingSantiago, LlipsyDomingo, María PilarRamírez-Labrada, ArielPaño, José RamónVendrell, MarcPardo, JuliánArias, MaykelGálvez Buerba, Eva MªGzmA activityProbeFluorescenceInflammatory diseasehttp://metadata.un.org/sdg/3Ensure healthy lives and promote well-being for all at all ages8 figures.-- Supplementary information available.Granzymes (Gzms), a family of serine proteases, expressed by immune and nonimmune cells, present perforin-dependent and independent intracellular and extracellular functions. When released in the extracellular space, GzmA, with trypsin-like activity, is involved in the pathophysiology of different inflammatory diseases. However, there are no validated specific systems to detect active forms of extracellular GzmA, making it difficult to assess its biological relevance and potential use as a biomarker. Here, we have developed fluorescence-energy resonance-transfer (FRET)-based peptide probes (FAM-peptide-DABCYL) to specifically detect GzmA activity in tissue samples and biological fluids in both mouse and human samples during inflammatory diseases. An initial probe was developed and incubated with GzmA and different proteases like GzmB and others with similar cleavage specificity as GzmA like GzmK, thrombin, trypsin, kallikrein, or plasmin. After measuring fluorescence, the probe showed very good specificity and sensitivity for human and mouse GzmA when compared to GzmB, its closest homologue GzmK, and with thrombin. The specificity of this probe was further refined by incubating the samples in a coated plate with a GzmA-specific antibody before adding the probe. The results show a high specific detection of soluble GzmA even when compared with other soluble proteases with very similar cleavage specificity like thrombin, GzmK, trypsin, kallikrein, or plasmin, which shows nearly no fluorescence signal. The high specific detection of GzmA was validated, showing that using pure proteins and serum and tissue samples from GzmA-deficient mice presented a significant reduction in the signal compared with WT mice. The utility of this system in humans was confirmed, showing that GzmA activity was significantly higher in serum samples from septic patients in comparison with healthy donors. Our results present a new immunoprobe with utility to detect extracellular GzmA activity in different biological fluids, confirming the presence of active forms of the soluble protease in vivo during inflammatory and infectious diseases.This research was supported by CIBER─Consorcio Centro de Investigación Biomédica en Red─CIBERINFEC (CB21-13-0087), Instituto de Salud Carlos III, Ministerio de Ciencia e Innovación and Unión Europea─NextGenerationEU, FEDER (Fondo Europeo de Desarrollo Regional), Gobierno de Aragón (Group B29_23R and LMP139_21), Ministerio de Ciencia, Innovación e Universidades (MCNU), Agencia Estatal de Investigación (PID2020-113963RB-I00, SAF2017-83120-C2-1-R). A.S.-S. was supported by a PhD fellowship from the Aragon Government. M.A. acknowledges funds from the Ministry of Economy and Competitiveness, Spain (FJCI-2017-31629, IJC2019-039192-I). M.V. acknowledges funds from an ERC Consolidator Grant (DYNAFLUORS, 771443), and this project has received funding from the European Union’s Horizon 2020 research and innovation program under the Marie Sklodowska-Curie grant agreement (859908).Peer reviewedACS PublicationsInstituto de Salud Carlos IIIMinisterio de Ciencia, Innovación y Universidades (España)European CommissionGobierno de AragónAgencia Estatal de Investigación (España)European Research CouncilMinisterio de Economía y Competitividad (España)Senán Salinas, Ana [0009-0009-5597-851X]Comas, Laura [0000-0002-3843-1231]Esteban, Patricia [0000-0003-4123-3524]Garzón, Marcela [0000-0001-6778-0636]Santiago, Llipsy [0000-0002-1861-5981]Domingo, María Pilar [0000-0002-6829-8769]Ramírez-Labrada, Ariel [0000-0002-3888-7036]Paño, José Ramón [0000-0002-9600-8116]Vendrell, Marc [0000-0002-5392-9740]Pardo, Julián [0000-0003-0154-0730]Arias, Maykel [0000-0002-9730-2210]Gálvez Buerba, Eva Mª [0000-0001-6928-5516]Gálvez Buerba, Eva Mª [eva@icb.csic.es]Arias, Maykel [maykelariascabrero@gmail.com]Consejo Superior de Investigaciones Científicas [https://ror.org/02gfc7t72]202420242024info:eu-repo/semantics/articlehttp://purl.org/coar/resource_type/c_6501Publisher's versioninfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10261/354919reponame:DIGITAL.CSIC. Repositorio Institucional del CSICinstname:Consejo Superior de Investigaciones Científicas (CSIC)Inglés#PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE##PLACEHOLDER_PARENT_METADATA_VALUE#info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2017-2020/PID2020-113963RB-I00info:eu-repo/grantAgreement/AEI/Plan Estatal de Investigación Científica y Técnica y de Innovación 2013-2016/SAF2017-83120-C2-1-Rinfo:eu-repo/grantAgreement/EC/H2020/771443info:eu-repo/grantAgreement/EC/H2020/859908Senán Salinas, Ana; Comas, Laura; Esteban, Patricia; Garzón, Marcela; Cheng, Zhiming; Santiago, Llipsy; Domingo, María Pilar; Ramírez-Labrada, Ariel; Paño, José Ramón; Vendrell, Marc; Pardo, Julián; Arias, Maykel; Gálvez Buerba, Eva Mª; 2024; Supporting information for Selective detection of active extracellular granzyme A by using a novel fluorescent immunoprobe with application to inflammatory diseases [Dataset]; ACS Publications; http://dx.doi.org/10.1021/acsptsci.4c00065https://www.wikidata.org/wiki/Q18026722http://dx.doi.org/10.1021/acsptsci.4c00065Síinfo:eu-repo/semantics/openAccessoai:digital.csic.es:10261/3549192026-05-22T06:33:51Z
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