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P2X-Receptor Antagonists Inhibit the Interaction of S. aureus Hemolysin A with Membranes

The pore forming hemolysin A, Hla, is a major virulence factor of Staphylococcus aureus. Apparently, 1–2 pore(s) per cell suffice(s) to cause cell death. Accumulated experimental evidence points towards a major role of ATP-gated purinergic receptors (P2XR) for hemolysis caused by Hla, complement and...

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Autores principales: Schwiering, Markus, Husmann, Matthias, Hellmann, Nadja
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5666378/
https://www.ncbi.nlm.nih.gov/pubmed/29048353
http://dx.doi.org/10.3390/toxins9100332
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author Schwiering, Markus
Husmann, Matthias
Hellmann, Nadja
author_facet Schwiering, Markus
Husmann, Matthias
Hellmann, Nadja
author_sort Schwiering, Markus
collection PubMed
description The pore forming hemolysin A, Hla, is a major virulence factor of Staphylococcus aureus. Apparently, 1–2 pore(s) per cell suffice(s) to cause cell death. Accumulated experimental evidence points towards a major role of ATP-gated purinergic receptors (P2XR) for hemolysis caused by Hla, complement and other pore forming proteins, presumably by increasing membrane permeability. Indeed, in experiments employing rabbit erythrocytes, inhibitory concentrations of frequently employed P2XR-antagonists were in a similar range as previously reported for erythrocytes of other species and other toxins. However, Hla-dependent hemolysis was not enhanced by extracellular ATP, and oxidized adenosinetriphosphate (oxATP) had only a minor inhibitory effect. Unexpectedly, P2XR-inhibitors also prevented Hla-induced lysis of pure lipid membranes, demonstrating that the inhibition did not even depend on the presence of P2XR. Fluorescence microscopy and gel-electrophoresis clearly revealed that P2XR-inhibitors interfere with binding and subsequent oligomerisation of Hla with membranes. Similar results were obtained employing HaCaT-cells. Furthermore, calorimetric data and hemolysis experiments with Hla pre-treated with pyridoxal phosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) showed that this compound directly binds to Hla. Our results call for a critical re-assessment of the appealing concept, which suggests that P2XR are general amplifiers of damage by pore-forming proteins.
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spelling pubmed-56663782017-11-09 P2X-Receptor Antagonists Inhibit the Interaction of S. aureus Hemolysin A with Membranes Schwiering, Markus Husmann, Matthias Hellmann, Nadja Toxins (Basel) Article The pore forming hemolysin A, Hla, is a major virulence factor of Staphylococcus aureus. Apparently, 1–2 pore(s) per cell suffice(s) to cause cell death. Accumulated experimental evidence points towards a major role of ATP-gated purinergic receptors (P2XR) for hemolysis caused by Hla, complement and other pore forming proteins, presumably by increasing membrane permeability. Indeed, in experiments employing rabbit erythrocytes, inhibitory concentrations of frequently employed P2XR-antagonists were in a similar range as previously reported for erythrocytes of other species and other toxins. However, Hla-dependent hemolysis was not enhanced by extracellular ATP, and oxidized adenosinetriphosphate (oxATP) had only a minor inhibitory effect. Unexpectedly, P2XR-inhibitors also prevented Hla-induced lysis of pure lipid membranes, demonstrating that the inhibition did not even depend on the presence of P2XR. Fluorescence microscopy and gel-electrophoresis clearly revealed that P2XR-inhibitors interfere with binding and subsequent oligomerisation of Hla with membranes. Similar results were obtained employing HaCaT-cells. Furthermore, calorimetric data and hemolysis experiments with Hla pre-treated with pyridoxal phosphate-6-azophenyl-2′,4′-disulfonic acid (PPADS) showed that this compound directly binds to Hla. Our results call for a critical re-assessment of the appealing concept, which suggests that P2XR are general amplifiers of damage by pore-forming proteins. MDPI 2017-10-19 /pmc/articles/PMC5666378/ /pubmed/29048353 http://dx.doi.org/10.3390/toxins9100332 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Schwiering, Markus
Husmann, Matthias
Hellmann, Nadja
P2X-Receptor Antagonists Inhibit the Interaction of S. aureus Hemolysin A with Membranes
title P2X-Receptor Antagonists Inhibit the Interaction of S. aureus Hemolysin A with Membranes
title_full P2X-Receptor Antagonists Inhibit the Interaction of S. aureus Hemolysin A with Membranes
title_fullStr P2X-Receptor Antagonists Inhibit the Interaction of S. aureus Hemolysin A with Membranes
title_full_unstemmed P2X-Receptor Antagonists Inhibit the Interaction of S. aureus Hemolysin A with Membranes
title_short P2X-Receptor Antagonists Inhibit the Interaction of S. aureus Hemolysin A with Membranes
title_sort p2x-receptor antagonists inhibit the interaction of s. aureus hemolysin a with membranes
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5666378/
https://www.ncbi.nlm.nih.gov/pubmed/29048353
http://dx.doi.org/10.3390/toxins9100332
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