Dynamic regulation of extracellular ATP in <i>Escherichia coli</i>

We studied the kinetics of extracellular ATP (ATPe) in Escherichia coli and their outer membrane vesicles (OMVs) stimulated with amphipatic peptides melittin (MEL) and mas-toparan 7 (MST7). Real-time luminometry was used to measure ATPe kinetics, ATP release, and ATPase activity. The latter was also...

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Autores principales: Alvarez, Cora Lilia, Corradi, Gerardo Raúl, Lauri, Natalia, Marginedas Freixa, Irene, Leal Denis, Maria Florencia, Enrique, Nicolás Jorge, Maté, Sabina María, Milesi, Verónica, Ostuni, Mariano Aníbal, Herlax, Vanesa Silvana, Schwarzbaum, Pablo Julio
Formato: Articulo
Lenguaje:Inglés
Publicado: 2017
Materias:
Ciencias Médicas
Caco-2 cells
Escherichia coli
Extracellular atp regulation
Mastoparan 7
Melittin
Outer membrane vesicles
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/96491
https://ri.conicet.gov.ar/11336/49158
http://www.biochemj.org/content/474/8/1395.long
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-96491
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Médicas
Caco-2 cells
Escherichia coli
Extracellular atp regulation
Mastoparan 7
Melittin
Outer membrane vesicles
spellingShingle Ciencias Médicas
Caco-2 cells
Escherichia coli
Extracellular atp regulation
Mastoparan 7
Melittin
Outer membrane vesicles
Alvarez, Cora Lilia
Corradi, Gerardo Raúl
Lauri, Natalia
Marginedas Freixa, Irene
Leal Denis, Maria Florencia
Enrique, Nicolás Jorge
Maté, Sabina María
Milesi, Verónica
Ostuni, Mariano Aníbal
Herlax, Vanesa Silvana
Schwarzbaum, Pablo Julio
Dynamic regulation of extracellular ATP in <i>Escherichia coli</i>
topic_facet Ciencias Médicas
Caco-2 cells
Escherichia coli
Extracellular atp regulation
Mastoparan 7
Melittin
Outer membrane vesicles
description We studied the kinetics of extracellular ATP (ATPe) in Escherichia coli and their outer membrane vesicles (OMVs) stimulated with amphipatic peptides melittin (MEL) and mas-toparan 7 (MST7). Real-time luminometry was used to measure ATPe kinetics, ATP release, and ATPase activity. The latter was also determined by following [32P]Pi released from [γ-32P]ATP. E. coli was studied alone, co-incubated with Caco-2 cells, or in rat jejunum segments. In E. coli, the addition of [γ-32P]ATP led to the uptake and subsequent hydrolysis of ATPe. Exposure to peptides caused an acute 3-fold (MST7) and 7-fold (MEL) increase in [ATPe]. In OMVs, ATPase activity increased linearly with [ATPe] (0.1-1 mM). Exposure to MST7 and MEL enhanced ATP release by 3-7 fold, with similar kinetics to that of bacteria. In Caco-2 cells, the addition of ATP to the apical domain led to a steep [ATPe] increase to a maximum, with subsequent ATPase activity. The addition of bacterial suspensions led to a 6-7 fold increase in [ATPe], followed by an acute decrease. In perfused jejunum segments, exposure to E. coli increased luminal ATP 2 fold. ATPe regulation of E. coli depends on the balance between ATPase activity and ATP release. This balance can be altered by OMVs, which display their own capacity to regulate ATPe. E. coli can activate ATP release from Caco-2 cells and intestinal segments, a response which in vivo might lead to intestinal release of ATP from the gut lumen.
format Articulo
Articulo
author Alvarez, Cora Lilia
Corradi, Gerardo Raúl
Lauri, Natalia
Marginedas Freixa, Irene
Leal Denis, Maria Florencia
Enrique, Nicolás Jorge
Maté, Sabina María
Milesi, Verónica
Ostuni, Mariano Aníbal
Herlax, Vanesa Silvana
Schwarzbaum, Pablo Julio
author_facet Alvarez, Cora Lilia
Corradi, Gerardo Raúl
Lauri, Natalia
Marginedas Freixa, Irene
Leal Denis, Maria Florencia
Enrique, Nicolás Jorge
Maté, Sabina María
Milesi, Verónica
Ostuni, Mariano Aníbal
Herlax, Vanesa Silvana
Schwarzbaum, Pablo Julio
author_sort Alvarez, Cora Lilia
title Dynamic regulation of extracellular ATP in <i>Escherichia coli</i>
title_short Dynamic regulation of extracellular ATP in <i>Escherichia coli</i>
title_full Dynamic regulation of extracellular ATP in <i>Escherichia coli</i>
title_fullStr Dynamic regulation of extracellular ATP in <i>Escherichia coli</i>
title_full_unstemmed Dynamic regulation of extracellular ATP in <i>Escherichia coli</i>
title_sort dynamic regulation of extracellular atp in <i>escherichia coli</i>
publishDate 2017
url http://sedici.unlp.edu.ar/handle/10915/96491
https://ri.conicet.gov.ar/11336/49158
http://www.biochemj.org/content/474/8/1395.long
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