An Improved Method for P2X7R Antagonist Screening

Detalhes bibliográficos
Autor(a) principal: Bezerra, Rômulo José Soares
Data de Publicação: 2015
Outros Autores: Ferreira, Natiele Carla da Silva, Alberto, Anael Viana Pinto, Binavita, André Gustavo, Fidalgo Neto, Antônio Augusto, Calgeiros, Andrea Surrage, Frutuoso, Valber da Silva, Alves, Luiz Anastácio
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/12780
Resumo: Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Comunicação Celular. Rio de Janeiro, RJ, Brasil.
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spelling Bezerra, Rômulo José SoaresFerreira, Natiele Carla da SilvaAlberto, Anael Viana PintoBinavita, André GustavoFidalgo Neto, Antônio AugustoCalgeiros, Andrea SurrageFrutuoso, Valber da SilvaAlves, Luiz Anastácio2016-02-22T16:12:42Z2016-02-22T16:12:42Z2015BEZERRA, Rômulo José Soares; et al. An Improved Method for P2X7R Antagonist Screening. Plos One, v.10, n.5, e0123089, 15p, May 2015.1932-6203https://www.arca.fiocruz.br/handle/icict/1278010.1371/journal.pone.0123089engPublic Library of ScienceAn Improved Method for P2X7R Antagonist Screeninginfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleFundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Comunicação Celular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Comunicação Celular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Comunicação Celular. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Laboratório de Pesquisa Integrada. Campus Macaé Professor Aloísio Teixeira. Macaé, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Comunicação Celular. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunofarmacologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Imunofarmacologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Comunicação Celular. Rio de Janeiro, RJ, Brasil.ATP physiologically activates the P2X7 receptor (P2X7R), a member of the P2X ionotropic receptor family. When activated by high concentrations of ATP (i.e., at inflammation sites), this receptor is capable of forming a pore that allows molecules of up to 900 Da to pass through. This receptor is upregulated in several diseases, particularly leukemia, rheumatoid arthritis and Alzheimer's disease. A selective antagonist of this receptor could be useful in the treatment of P2X7R activation-related diseases. In the present study, we have evaluated several parameters using in vitro protocols to validate a high-throughput screening (HTS) method to identify P2X7R antagonists. We generated dose-response curves to determine the EC50 value of the known agonist ATP and the ICs50 values for the known antagonists Brilliant Blue G (BBG) and oxidized ATP (OATP). The values obtained were consistent with those found in the literature (0.7 ± 0.07 mM, 1.3-2.6 mM and 173-285 μM for ATP, BBG and OATP, respectively). The Z-factor, an important statistical tool that can be used to validate the robustness and suitability of an HTS assay, was 0.635 for PI uptake and 0.867 for LY uptake. No inter-operator variation was observed, and the results obtained using our improved method were reproducible. Our data indicate that our assay is suitable for the selective and reliable evaluation of P2X7 activity in multiwell plates using spectrophotometry-based methodology. This method might improve the high-throughput screening of conventional chemical or natural product libraries for possible candidate P2X7R antagonist or agonist.P2X7 activitySpectrofluorometryP2X7R Antagonist ScreeningP2X ionotropic receptor familyStructure-Activity Relationshipinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-82991https://www.arca.fiocruz.br/bitstream/icict/12780/1/license.txt5a560609d32a3863062d77ff32785d58MD51ORIGINALromulo_bezerra_etal_IOC_2015.pdfromulo_bezerra_etal_IOC_2015.pdfapplication/pdf3138870https://www.arca.fiocruz.br/bitstream/icict/12780/2/romulo_bezerra_etal_IOC_2015.pdfc870dbe9ab0a1619cb03088d944f371fMD52TEXTromulo_bezerra_etal_IOC_2015.pdf.txtromulo_bezerra_etal_IOC_2015.pdf.txtExtracted 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dc.title.pt_BR.fl_str_mv An Improved Method for P2X7R Antagonist Screening
title An Improved Method for P2X7R Antagonist Screening
spellingShingle An Improved Method for P2X7R Antagonist Screening
Bezerra, Rômulo José Soares
P2X7 activity
Spectrofluorometry
P2X7R Antagonist Screening
P2X ionotropic receptor family
Structure-Activity Relationship
title_short An Improved Method for P2X7R Antagonist Screening
title_full An Improved Method for P2X7R Antagonist Screening
title_fullStr An Improved Method for P2X7R Antagonist Screening
title_full_unstemmed An Improved Method for P2X7R Antagonist Screening
title_sort An Improved Method for P2X7R Antagonist Screening
author Bezerra, Rômulo José Soares
author_facet Bezerra, Rômulo José Soares
Ferreira, Natiele Carla da Silva
Alberto, Anael Viana Pinto
Binavita, André Gustavo
Fidalgo Neto, Antônio Augusto
Calgeiros, Andrea Surrage
Frutuoso, Valber da Silva
Alves, Luiz Anastácio
author_role author
author2 Ferreira, Natiele Carla da Silva
Alberto, Anael Viana Pinto
Binavita, André Gustavo
Fidalgo Neto, Antônio Augusto
Calgeiros, Andrea Surrage
Frutuoso, Valber da Silva
Alves, Luiz Anastácio
author2_role author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Bezerra, Rômulo José Soares
Ferreira, Natiele Carla da Silva
Alberto, Anael Viana Pinto
Binavita, André Gustavo
Fidalgo Neto, Antônio Augusto
Calgeiros, Andrea Surrage
Frutuoso, Valber da Silva
Alves, Luiz Anastácio
dc.subject.en.pt_BR.fl_str_mv P2X7 activity
Spectrofluorometry
P2X7R Antagonist Screening
P2X ionotropic receptor family
Structure-Activity Relationship
topic P2X7 activity
Spectrofluorometry
P2X7R Antagonist Screening
P2X ionotropic receptor family
Structure-Activity Relationship
description Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Laboratório de Comunicação Celular. Rio de Janeiro, RJ, Brasil.
publishDate 2015
dc.date.issued.fl_str_mv 2015
dc.date.accessioned.fl_str_mv 2016-02-22T16:12:42Z
dc.date.available.fl_str_mv 2016-02-22T16:12:42Z
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.citation.fl_str_mv BEZERRA, Rômulo José Soares; et al. An Improved Method for P2X7R Antagonist Screening. Plos One, v.10, n.5, e0123089, 15p, May 2015.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/12780
dc.identifier.issn.none.fl_str_mv 1932-6203
dc.identifier.doi.none.fl_str_mv 10.1371/journal.pone.0123089
identifier_str_mv BEZERRA, Rômulo José Soares; et al. An Improved Method for P2X7R Antagonist Screening. Plos One, v.10, n.5, e0123089, 15p, May 2015.
1932-6203
10.1371/journal.pone.0123089
url https://www.arca.fiocruz.br/handle/icict/12780
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Public Library of Science
publisher.none.fl_str_mv Public Library of Science
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
instname_str Fundação Oswaldo Cruz (FIOCRUZ)
instacron_str FIOCRUZ
institution FIOCRUZ
reponame_str Repositório Institucional da FIOCRUZ (ARCA)
collection Repositório Institucional da FIOCRUZ (ARCA)
bitstream.url.fl_str_mv https://www.arca.fiocruz.br/bitstream/icict/12780/1/license.txt
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