Staphylococcal Pore-forming Leukotoxins: Opening of Ca<sup>2+</sup>-activated K<sup>+</sup> Channels and Specificity of Membrane Pores in Human Neutrophils

Leila Staali; Didier Andre Colin

doi:doi:10.11648/j.ijmb.20200503.19

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Staphylococcal Pore-forming Leukotoxins: Opening of Ca²⁺-activated K⁺ Channels and Specificity of Membrane Pores in Human Neutrophils

Leila Staali, Didier Andre Colin

Published in International Journal of Microbiology and Biotechnology (Volume 5, Issue 3)

Received: 14 July 2020 Accepted: 27 July 2020 Published: 20 August 2020

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Abstract

Pore-forming toxins are key virulence determinants produced by human bacterial pathogens Staphylococcus aureus, inducing two independent cellular events in neutrophils. Upon a specific binding to membrane receptors, both Panton and Valentin Leukocidin and γ-hemolysin induced an increase of Na⁺ and K⁺ fluxes, likely associated to the activation of preexisting ionic channels or to the membrane pores formation. This was investigated by using, spectrofluorometry techniques and, specific molecular probes in human neutrophils. Interestingly, we found that, in the absence of extracellular Ca²⁺, leukotoxins did form membrane pores, which were large enough to allow a massive entry of ethidium into neutrophils. Simultaneously, sustained Na⁺ influx and K⁺ efflux were observed. Another set of experiments carried out in the presence of extracellular Ca²⁺ did show that, the percentage of pores formed by leukotoxins was significantly, reduced due to the Ca²⁺ effect to eventually protect cells from lysis. The simultaneous recording of Na⁺ and K⁺ movements showed a significant increase of the K⁺ efflux although, the Na⁺ influx was reduced. By using potassium channels blockers, we found that, the potassium efflux enhanced by the presence of extracellular Ca²⁺, was markedly, inhibited in apamin-, charybdotoxin-, tetrodotoxin-, and quinine-pretreatment neutrophils. We also found that, the increase of the K⁺ efflux was reduced by either, thapsigargin or TMB8, potent blockers of the internal Ca²⁺ stores depletion. Consequently, we proposed that, the activation of another potassium pathway by leukotoxins, known as Ca²⁺-activated K⁺ channels following the Ca²⁺ stores depletion. Furthermore, potassium channels blockers did not affect ethidium, Na⁺ and K⁺ movements, in the absence of extracellular Ca²⁺. Moreover, in this condition, no monovalent ions movement was recorded, when the pores formation was altered by tetra-ethyl-ammonium. In the present study, we further highlighted the specificity of membrane pores to Na⁺ and K⁺ ions when, the pores formation was completely blocked by divalent ions blockers (Ca²⁺ and Zn²⁺). Under these conditions, no monovalent ions movement, was recorded although, a significant influx of Ca²⁺ and Zn²⁺ was observed after the leukotoxins application. In conclusion, our data provided an evidence that, staphylococcal leukotoxins induced in human neutrophils: 1) the opening of Ca²⁺-activated K⁺ channels, only in the presence of 1 mM extracellular Ca²⁺; 2) the formation of membrane pores, which exhibited a high specificity to monovalent cations and, 3) an influx of sodium, through a tetrodotoxin not-sensitive pathway ruling out the hypothesis that, Na⁺ channels could be activated by leukotoxins.

Published in	International Journal of Microbiology and Biotechnology (Volume 5, Issue 3)
DOI	10.11648/j.ijmb.20200503.19
Page(s)	135-151
Creative Commons	This is an Open Access article, distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium or format, provided the original work is properly cited.
Copyright	Copyright © The Author(s), 2020. Published by Science Publishing Group

Keywords

Staphylococcal Pore-forming Leukotoxins: Opening of Ca²⁺-activated K⁺ Channels and Specificity of Membrane Pores in Human Neutrophils

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Leila Staali, Didier Andre Colin. (2020). Staphylococcal Pore-forming Leukotoxins: Opening of Ca2+-activated K+ Channels and Specificity of Membrane Pores in Human Neutrophils. International Journal of Microbiology and Biotechnology, 5(3), 135-151. https://doi.org/10.11648/j.ijmb.20200503.19

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Leila Staali; Didier Andre Colin. Staphylococcal Pore-forming Leukotoxins: Opening of Ca2+-activated K+ Channels and Specificity of Membrane Pores in Human Neutrophils. Int. J. Microbiol. Biotechnol. 2020, 5(3), 135-151. doi: 10.11648/j.ijmb.20200503.19

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Leila Staali, Didier Andre Colin. Staphylococcal Pore-forming Leukotoxins: Opening of Ca2+-activated K+ Channels and Specificity of Membrane Pores in Human Neutrophils. Int J Microbiol Biotechnol. 2020;5(3):135-151. doi: 10.11648/j.ijmb.20200503.19

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@article{10.11648/j.ijmb.20200503.19,
  author = {Leila Staali and Didier Andre Colin},
  title = {Staphylococcal Pore-forming Leukotoxins: Opening of Ca2+-activated K+ Channels and Specificity of Membrane Pores in Human Neutrophils},
  journal = {International Journal of Microbiology and Biotechnology},
  volume = {5},
  number = {3},
  pages = {135-151},
  doi = {10.11648/j.ijmb.20200503.19},
  url = {https://doi.org/10.11648/j.ijmb.20200503.19},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20200503.19},
  abstract = {Pore-forming toxins are key virulence determinants produced by human bacterial pathogens Staphylococcus aureus, inducing two independent cellular events in neutrophils. Upon a specific binding to membrane receptors, both Panton and Valentin Leukocidin and γ-hemolysin induced an increase of Na+ and K+ fluxes, likely associated to the activation of preexisting ionic channels or to the membrane pores formation. This was investigated by using, spectrofluorometry techniques and, specific molecular probes in human neutrophils. Interestingly, we found that, in the absence of extracellular Ca2+, leukotoxins did form membrane pores, which were large enough to allow a massive entry of ethidium into neutrophils. Simultaneously, sustained Na+ influx and K+ efflux were observed. Another set of experiments carried out in the presence of extracellular Ca2+ did show that, the percentage of pores formed by leukotoxins was significantly, reduced due to the Ca2+ effect to eventually protect cells from lysis. The simultaneous recording of Na+ and K+ movements showed a significant increase of the K+ efflux although, the Na+ influx was reduced. By using potassium channels blockers, we found that, the potassium efflux enhanced by the presence of extracellular Ca2+, was markedly, inhibited in apamin-, charybdotoxin-, tetrodotoxin-, and quinine-pretreatment neutrophils. We also found that, the increase of the K+ efflux was reduced by either, thapsigargin or TMB8, potent blockers of the internal Ca2+ stores depletion. Consequently, we proposed that, the activation of another potassium pathway by leukotoxins, known as Ca2+-activated K+ channels following the Ca2+ stores depletion. Furthermore, potassium channels blockers did not affect ethidium, Na+ and K+ movements, in the absence of extracellular Ca2+. Moreover, in this condition, no monovalent ions movement was recorded, when the pores formation was altered by tetra-ethyl-ammonium. In the present study, we further highlighted the specificity of membrane pores to Na+ and K+ ions when, the pores formation was completely blocked by divalent ions blockers (Ca2+ and Zn2+). Under these conditions, no monovalent ions movement, was recorded although, a significant influx of Ca2+ and Zn2+ was observed after the leukotoxins application. In conclusion, our data provided an evidence that, staphylococcal leukotoxins induced in human neutrophils: 1) the opening of Ca2+-activated K+ channels, only in the presence of 1 mM extracellular Ca2+; 2) the formation of membrane pores, which exhibited a high specificity to monovalent cations and, 3) an influx of sodium, through a tetrodotoxin not-sensitive pathway ruling out the hypothesis that, Na+ channels could be activated by leukotoxins.},
 year = {2020}
}

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TY  - JOUR
T1  - Staphylococcal Pore-forming Leukotoxins: Opening of Ca2+-activated K+ Channels and Specificity of Membrane Pores in Human Neutrophils
AU  - Leila Staali
AU  - Didier Andre Colin
Y1  - 2020/08/20
PY  - 2020
N1  - https://doi.org/10.11648/j.ijmb.20200503.19
DO  - 10.11648/j.ijmb.20200503.19
T2  - International Journal of Microbiology and Biotechnology
JF  - International Journal of Microbiology and Biotechnology
JO  - International Journal of Microbiology and Biotechnology
SP  - 135
EP  - 151
PB  - Science Publishing Group
SN  - 2578-9686
UR  - https://doi.org/10.11648/j.ijmb.20200503.19
AB  - Pore-forming toxins are key virulence determinants produced by human bacterial pathogens Staphylococcus aureus, inducing two independent cellular events in neutrophils. Upon a specific binding to membrane receptors, both Panton and Valentin Leukocidin and γ-hemolysin induced an increase of Na+ and K+ fluxes, likely associated to the activation of preexisting ionic channels or to the membrane pores formation. This was investigated by using, spectrofluorometry techniques and, specific molecular probes in human neutrophils. Interestingly, we found that, in the absence of extracellular Ca2+, leukotoxins did form membrane pores, which were large enough to allow a massive entry of ethidium into neutrophils. Simultaneously, sustained Na+ influx and K+ efflux were observed. Another set of experiments carried out in the presence of extracellular Ca2+ did show that, the percentage of pores formed by leukotoxins was significantly, reduced due to the Ca2+ effect to eventually protect cells from lysis. The simultaneous recording of Na+ and K+ movements showed a significant increase of the K+ efflux although, the Na+ influx was reduced. By using potassium channels blockers, we found that, the potassium efflux enhanced by the presence of extracellular Ca2+, was markedly, inhibited in apamin-, charybdotoxin-, tetrodotoxin-, and quinine-pretreatment neutrophils. We also found that, the increase of the K+ efflux was reduced by either, thapsigargin or TMB8, potent blockers of the internal Ca2+ stores depletion. Consequently, we proposed that, the activation of another potassium pathway by leukotoxins, known as Ca2+-activated K+ channels following the Ca2+ stores depletion. Furthermore, potassium channels blockers did not affect ethidium, Na+ and K+ movements, in the absence of extracellular Ca2+. Moreover, in this condition, no monovalent ions movement was recorded, when the pores formation was altered by tetra-ethyl-ammonium. In the present study, we further highlighted the specificity of membrane pores to Na+ and K+ ions when, the pores formation was completely blocked by divalent ions blockers (Ca2+ and Zn2+). Under these conditions, no monovalent ions movement, was recorded although, a significant influx of Ca2+ and Zn2+ was observed after the leukotoxins application. In conclusion, our data provided an evidence that, staphylococcal leukotoxins induced in human neutrophils: 1) the opening of Ca2+-activated K+ channels, only in the presence of 1 mM extracellular Ca2+; 2) the formation of membrane pores, which exhibited a high specificity to monovalent cations and, 3) an influx of sodium, through a tetrodotoxin not-sensitive pathway ruling out the hypothesis that, Na+ channels could be activated by leukotoxins.
VL  - 5
IS  - 3
ER  -

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Author Information

Leila Staali

Department of Biotechnology, Natural and Life Sciences Faculty, Ahmed Ben Bella Oran1-University, Oran, Algeria
Didier Andre Colin

Bacteriology Institute of Medical Faculty, Louis Pasteur University, Strasbourg, France

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APA Style

Leila Staali, Didier Andre Colin. (2020). Staphylococcal Pore-forming Leukotoxins: Opening of Ca2+-activated K+ Channels and Specificity of Membrane Pores in Human Neutrophils. International Journal of Microbiology and Biotechnology, 5(3), 135-151. https://doi.org/10.11648/j.ijmb.20200503.19

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ACS Style

Leila Staali; Didier Andre Colin. Staphylococcal Pore-forming Leukotoxins: Opening of Ca2+-activated K+ Channels and Specificity of Membrane Pores in Human Neutrophils. Int. J. Microbiol. Biotechnol. 2020, 5(3), 135-151. doi: 10.11648/j.ijmb.20200503.19

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AMA Style

Leila Staali, Didier Andre Colin. Staphylococcal Pore-forming Leukotoxins: Opening of Ca2+-activated K+ Channels and Specificity of Membrane Pores in Human Neutrophils. Int J Microbiol Biotechnol. 2020;5(3):135-151. doi: 10.11648/j.ijmb.20200503.19

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@article{10.11648/j.ijmb.20200503.19,
  author = {Leila Staali and Didier Andre Colin},
  title = {Staphylococcal Pore-forming Leukotoxins: Opening of Ca2+-activated K+ Channels and Specificity of Membrane Pores in Human Neutrophils},
  journal = {International Journal of Microbiology and Biotechnology},
  volume = {5},
  number = {3},
  pages = {135-151},
  doi = {10.11648/j.ijmb.20200503.19},
  url = {https://doi.org/10.11648/j.ijmb.20200503.19},
  eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20200503.19},
  abstract = {Pore-forming toxins are key virulence determinants produced by human bacterial pathogens Staphylococcus aureus, inducing two independent cellular events in neutrophils. Upon a specific binding to membrane receptors, both Panton and Valentin Leukocidin and γ-hemolysin induced an increase of Na+ and K+ fluxes, likely associated to the activation of preexisting ionic channels or to the membrane pores formation. This was investigated by using, spectrofluorometry techniques and, specific molecular probes in human neutrophils. Interestingly, we found that, in the absence of extracellular Ca2+, leukotoxins did form membrane pores, which were large enough to allow a massive entry of ethidium into neutrophils. Simultaneously, sustained Na+ influx and K+ efflux were observed. Another set of experiments carried out in the presence of extracellular Ca2+ did show that, the percentage of pores formed by leukotoxins was significantly, reduced due to the Ca2+ effect to eventually protect cells from lysis. The simultaneous recording of Na+ and K+ movements showed a significant increase of the K+ efflux although, the Na+ influx was reduced. By using potassium channels blockers, we found that, the potassium efflux enhanced by the presence of extracellular Ca2+, was markedly, inhibited in apamin-, charybdotoxin-, tetrodotoxin-, and quinine-pretreatment neutrophils. We also found that, the increase of the K+ efflux was reduced by either, thapsigargin or TMB8, potent blockers of the internal Ca2+ stores depletion. Consequently, we proposed that, the activation of another potassium pathway by leukotoxins, known as Ca2+-activated K+ channels following the Ca2+ stores depletion. Furthermore, potassium channels blockers did not affect ethidium, Na+ and K+ movements, in the absence of extracellular Ca2+. Moreover, in this condition, no monovalent ions movement was recorded, when the pores formation was altered by tetra-ethyl-ammonium. In the present study, we further highlighted the specificity of membrane pores to Na+ and K+ ions when, the pores formation was completely blocked by divalent ions blockers (Ca2+ and Zn2+). Under these conditions, no monovalent ions movement, was recorded although, a significant influx of Ca2+ and Zn2+ was observed after the leukotoxins application. In conclusion, our data provided an evidence that, staphylococcal leukotoxins induced in human neutrophils: 1) the opening of Ca2+-activated K+ channels, only in the presence of 1 mM extracellular Ca2+; 2) the formation of membrane pores, which exhibited a high specificity to monovalent cations and, 3) an influx of sodium, through a tetrodotoxin not-sensitive pathway ruling out the hypothesis that, Na+ channels could be activated by leukotoxins.},
 year = {2020}
}

Copy | Download

TY  - JOUR
T1  - Staphylococcal Pore-forming Leukotoxins: Opening of Ca2+-activated K+ Channels and Specificity of Membrane Pores in Human Neutrophils
AU  - Leila Staali
AU  - Didier Andre Colin
Y1  - 2020/08/20
PY  - 2020
N1  - https://doi.org/10.11648/j.ijmb.20200503.19
DO  - 10.11648/j.ijmb.20200503.19
T2  - International Journal of Microbiology and Biotechnology
JF  - International Journal of Microbiology and Biotechnology
JO  - International Journal of Microbiology and Biotechnology
SP  - 135
EP  - 151
PB  - Science Publishing Group
SN  - 2578-9686
UR  - https://doi.org/10.11648/j.ijmb.20200503.19
AB  - Pore-forming toxins are key virulence determinants produced by human bacterial pathogens Staphylococcus aureus, inducing two independent cellular events in neutrophils. Upon a specific binding to membrane receptors, both Panton and Valentin Leukocidin and γ-hemolysin induced an increase of Na+ and K+ fluxes, likely associated to the activation of preexisting ionic channels or to the membrane pores formation. This was investigated by using, spectrofluorometry techniques and, specific molecular probes in human neutrophils. Interestingly, we found that, in the absence of extracellular Ca2+, leukotoxins did form membrane pores, which were large enough to allow a massive entry of ethidium into neutrophils. Simultaneously, sustained Na+ influx and K+ efflux were observed. Another set of experiments carried out in the presence of extracellular Ca2+ did show that, the percentage of pores formed by leukotoxins was significantly, reduced due to the Ca2+ effect to eventually protect cells from lysis. The simultaneous recording of Na+ and K+ movements showed a significant increase of the K+ efflux although, the Na+ influx was reduced. By using potassium channels blockers, we found that, the potassium efflux enhanced by the presence of extracellular Ca2+, was markedly, inhibited in apamin-, charybdotoxin-, tetrodotoxin-, and quinine-pretreatment neutrophils. We also found that, the increase of the K+ efflux was reduced by either, thapsigargin or TMB8, potent blockers of the internal Ca2+ stores depletion. Consequently, we proposed that, the activation of another potassium pathway by leukotoxins, known as Ca2+-activated K+ channels following the Ca2+ stores depletion. Furthermore, potassium channels blockers did not affect ethidium, Na+ and K+ movements, in the absence of extracellular Ca2+. Moreover, in this condition, no monovalent ions movement was recorded, when the pores formation was altered by tetra-ethyl-ammonium. In the present study, we further highlighted the specificity of membrane pores to Na+ and K+ ions when, the pores formation was completely blocked by divalent ions blockers (Ca2+ and Zn2+). Under these conditions, no monovalent ions movement, was recorded although, a significant influx of Ca2+ and Zn2+ was observed after the leukotoxins application. In conclusion, our data provided an evidence that, staphylococcal leukotoxins induced in human neutrophils: 1) the opening of Ca2+-activated K+ channels, only in the presence of 1 mM extracellular Ca2+; 2) the formation of membrane pores, which exhibited a high specificity to monovalent cations and, 3) an influx of sodium, through a tetrodotoxin not-sensitive pathway ruling out the hypothesis that, Na+ channels could be activated by leukotoxins.
VL  - 5
IS  - 3
ER  -

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