pH Modulation of efflux pump activity of multi-drug resistant Escherichia coli protection during its passage and eventual colonization of the colon
Autor(a) principal: | |
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Data de Publicação: | 2009 |
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/10362/117219 |
Resumo: | BACKGROUND: Resistance Nodulation Division (RND) efflux pumps of Escherichia coli extrude antibiotics and toxic substances before they reach their intended targets. Whereas these pumps obtain their energy directly from the proton motive force (PMF), ATP-Binding Cassette (ABC) transporters, which can also extrude antibiotics, obtain energy from the hydrolysis of ATP. Because E. coli must pass through two pH distinct environments of the gastrointestinal system of the host, it must be able to extrude toxic agents at very acidic and at near neutral pH (bile salts in duodenum and colon for example). The herein described study examines the effect of pH on the extrusion of ethidium bromide (EB). METHODOLOGY/PRINCIPAL FINDINGS: E. coli AG100 and its tetracycline induced progeny AG100(TET) that over-expresses the acrAB efflux pump were evaluated for their ability to extrude EB at pH 5 and 8, by our recently developed semi-automated fluorometric method. At pH 5 the organism extrudes EB without the need for metabolic energy (glucose), whereas at pH 8 extrusion of EB is dependent upon metabolic energy. Phe-Arg beta-naphtylamide (PAbetaN), a commonly assumed inhibitor of RND efflux pumps has no effect on the extrusion of EB as others claim. However, it does cause accumulation of EB. Competition between EB and PAbetaN was demonstrated and suggested that PAbetaN was preferentially extruded. A K(m) representing competition between PAbetaN and EB has been calculated. CONCLUSIONS/SIGNIFICANCE: The results suggest that E. coli has two general efflux systems (not to be confused with a distinct efflux pump) that are activated at low and high pH, respectively, and that the one at high pH is probably a putative ABC transporter coded by msbA, which has significant homology to the ABC transporter coded by efrAB of Enterococcus faecalis, an organism that faces similar challenges as it makes its way through the toxic intestinal system of the host. |
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pH Modulation of efflux pump activity of multi-drug resistant Escherichia coli protection during its passage and eventual colonization of the colonBiological TransportColonDrug Resistance, MultipleEscherichia coliHydrogen-Ion ConcentrationMicrobial Sensitivity TestsJournal ArticleResearch Support, Non-U.S. Gov'tMicrobiologyBiochemistry, Genetics and Molecular Biology (miscellaneous)Infectious DiseasesSDG 3 - Good Health and Well-beingBACKGROUND: Resistance Nodulation Division (RND) efflux pumps of Escherichia coli extrude antibiotics and toxic substances before they reach their intended targets. Whereas these pumps obtain their energy directly from the proton motive force (PMF), ATP-Binding Cassette (ABC) transporters, which can also extrude antibiotics, obtain energy from the hydrolysis of ATP. Because E. coli must pass through two pH distinct environments of the gastrointestinal system of the host, it must be able to extrude toxic agents at very acidic and at near neutral pH (bile salts in duodenum and colon for example). The herein described study examines the effect of pH on the extrusion of ethidium bromide (EB). METHODOLOGY/PRINCIPAL FINDINGS: E. coli AG100 and its tetracycline induced progeny AG100(TET) that over-expresses the acrAB efflux pump were evaluated for their ability to extrude EB at pH 5 and 8, by our recently developed semi-automated fluorometric method. At pH 5 the organism extrudes EB without the need for metabolic energy (glucose), whereas at pH 8 extrusion of EB is dependent upon metabolic energy. Phe-Arg beta-naphtylamide (PAbetaN), a commonly assumed inhibitor of RND efflux pumps has no effect on the extrusion of EB as others claim. However, it does cause accumulation of EB. Competition between EB and PAbetaN was demonstrated and suggested that PAbetaN was preferentially extruded. A K(m) representing competition between PAbetaN and EB has been calculated. CONCLUSIONS/SIGNIFICANCE: The results suggest that E. coli has two general efflux systems (not to be confused with a distinct efflux pump) that are activated at low and high pH, respectively, and that the one at high pH is probably a putative ABC transporter coded by msbA, which has significant homology to the ABC transporter coded by efrAB of Enterococcus faecalis, an organism that faces similar challenges as it makes its way through the toxic intestinal system of the host.Unidade de Parasitologia e Microbiologia Médicas (UPMM)Instituto de Higiene e Medicina Tropical (IHMT)CREM - Centro de Recursos MicrobiológicosRUNMartins, AnaSpengler, GabriellaRodrigues, LilianaViveiros, MiguelRamos, JorgeMartins, MartaCouto, IsabelFanning, SéamusPagès, Jean MarieBolla, Jean MichelMolnar, JosephAmaral, Leonard2021-05-06T22:40:05Z2009-08-172009-08-17T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article9application/pdfhttp://hdl.handle.net/10362/117219eng1932-6203PURE: 2579292https://doi.org/10.1371/journal.pone.0006656info: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:RCAAP2024-03-11T05:00:19Zoai:run.unl.pt:10362/117219Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T03:43:32.584858Repositório 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 |
pH Modulation of efflux pump activity of multi-drug resistant Escherichia coli protection during its passage and eventual colonization of the colon |
title |
pH Modulation of efflux pump activity of multi-drug resistant Escherichia coli protection during its passage and eventual colonization of the colon |
spellingShingle |
pH Modulation of efflux pump activity of multi-drug resistant Escherichia coli protection during its passage and eventual colonization of the colon Martins, Ana Biological Transport Colon Drug Resistance, Multiple Escherichia coli Hydrogen-Ion Concentration Microbial Sensitivity Tests Journal Article Research Support, Non-U.S. Gov't Microbiology Biochemistry, Genetics and Molecular Biology (miscellaneous) Infectious Diseases SDG 3 - Good Health and Well-being |
title_short |
pH Modulation of efflux pump activity of multi-drug resistant Escherichia coli protection during its passage and eventual colonization of the colon |
title_full |
pH Modulation of efflux pump activity of multi-drug resistant Escherichia coli protection during its passage and eventual colonization of the colon |
title_fullStr |
pH Modulation of efflux pump activity of multi-drug resistant Escherichia coli protection during its passage and eventual colonization of the colon |
title_full_unstemmed |
pH Modulation of efflux pump activity of multi-drug resistant Escherichia coli protection during its passage and eventual colonization of the colon |
title_sort |
pH Modulation of efflux pump activity of multi-drug resistant Escherichia coli protection during its passage and eventual colonization of the colon |
author |
Martins, Ana |
author_facet |
Martins, Ana Spengler, Gabriella Rodrigues, Liliana Viveiros, Miguel Ramos, Jorge Martins, Marta Couto, Isabel Fanning, Séamus Pagès, Jean Marie Bolla, Jean Michel Molnar, Joseph Amaral, Leonard |
author_role |
author |
author2 |
Spengler, Gabriella Rodrigues, Liliana Viveiros, Miguel Ramos, Jorge Martins, Marta Couto, Isabel Fanning, Séamus Pagès, Jean Marie Bolla, Jean Michel Molnar, Joseph Amaral, Leonard |
author2_role |
author author author author author author author author author author author |
dc.contributor.none.fl_str_mv |
Unidade de Parasitologia e Microbiologia Médicas (UPMM) Instituto de Higiene e Medicina Tropical (IHMT) CREM - Centro de Recursos Microbiológicos RUN |
dc.contributor.author.fl_str_mv |
Martins, Ana Spengler, Gabriella Rodrigues, Liliana Viveiros, Miguel Ramos, Jorge Martins, Marta Couto, Isabel Fanning, Séamus Pagès, Jean Marie Bolla, Jean Michel Molnar, Joseph Amaral, Leonard |
dc.subject.por.fl_str_mv |
Biological Transport Colon Drug Resistance, Multiple Escherichia coli Hydrogen-Ion Concentration Microbial Sensitivity Tests Journal Article Research Support, Non-U.S. Gov't Microbiology Biochemistry, Genetics and Molecular Biology (miscellaneous) Infectious Diseases SDG 3 - Good Health and Well-being |
topic |
Biological Transport Colon Drug Resistance, Multiple Escherichia coli Hydrogen-Ion Concentration Microbial Sensitivity Tests Journal Article Research Support, Non-U.S. Gov't Microbiology Biochemistry, Genetics and Molecular Biology (miscellaneous) Infectious Diseases SDG 3 - Good Health and Well-being |
description |
BACKGROUND: Resistance Nodulation Division (RND) efflux pumps of Escherichia coli extrude antibiotics and toxic substances before they reach their intended targets. Whereas these pumps obtain their energy directly from the proton motive force (PMF), ATP-Binding Cassette (ABC) transporters, which can also extrude antibiotics, obtain energy from the hydrolysis of ATP. Because E. coli must pass through two pH distinct environments of the gastrointestinal system of the host, it must be able to extrude toxic agents at very acidic and at near neutral pH (bile salts in duodenum and colon for example). The herein described study examines the effect of pH on the extrusion of ethidium bromide (EB). METHODOLOGY/PRINCIPAL FINDINGS: E. coli AG100 and its tetracycline induced progeny AG100(TET) that over-expresses the acrAB efflux pump were evaluated for their ability to extrude EB at pH 5 and 8, by our recently developed semi-automated fluorometric method. At pH 5 the organism extrudes EB without the need for metabolic energy (glucose), whereas at pH 8 extrusion of EB is dependent upon metabolic energy. Phe-Arg beta-naphtylamide (PAbetaN), a commonly assumed inhibitor of RND efflux pumps has no effect on the extrusion of EB as others claim. However, it does cause accumulation of EB. Competition between EB and PAbetaN was demonstrated and suggested that PAbetaN was preferentially extruded. A K(m) representing competition between PAbetaN and EB has been calculated. CONCLUSIONS/SIGNIFICANCE: The results suggest that E. coli has two general efflux systems (not to be confused with a distinct efflux pump) that are activated at low and high pH, respectively, and that the one at high pH is probably a putative ABC transporter coded by msbA, which has significant homology to the ABC transporter coded by efrAB of Enterococcus faecalis, an organism that faces similar challenges as it makes its way through the toxic intestinal system of the host. |
publishDate |
2009 |
dc.date.none.fl_str_mv |
2009-08-17 2009-08-17T00:00:00Z 2021-05-06T22:40:05Z |
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/10362/117219 |
url |
http://hdl.handle.net/10362/117219 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
1932-6203 PURE: 2579292 https://doi.org/10.1371/journal.pone.0006656 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
9 application/pdf |
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) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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