Mechanism of ligand-gated potassium efflux in bacterial pathogens

Tarmo P Roosild (Corresponding Author), Samantha Castronovo, Jess Healy, Samantha Miller, Christos Pliotas, Tim Rasmussen, Wendy Bartlett, Stuart J Conway, Ian R Booth (Corresponding Author)

Research output: Contribution to journalArticle

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Abstract

Gram negative pathogens are protected against toxic electrophilic compounds by glutathione-gated potassium efflux systems (Kef) that modulate cytoplasmic pH. We have elucidated the mechanism of gating through structural and functional analysis of Escherichia coli KefC. The revealed mechanism can explain how subtle chemical differences in glutathione derivatives can produce opposite effects on channel function. Kef channels are regulated by potassium transport and NAD-binding (KTN) domains that sense both reduced glutathione, which inhibits Kef activity, and glutathione adducts that form during electrophile detoxification and activate Kef. We find that reduced glutathione stabilizes an interdomain association between two KTN folds, whereas large adducts sterically disrupt this interaction. F441 is identified as the pivotal residue discriminating between reduced glutathione and its conjugates. We demonstrate a major structural change on the binding of an activating ligand to a KTN-domain protein. Analysis of the regulatory interactions suggests strategies to disrupt pathogen potassium and pH homeostasis.
Original languageEnglish
Pages (from-to)19784-19789
Number of pages6
JournalPNAS
Volume107
Issue number46
DOIs
Publication statusPublished - 16 Nov 2010

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Glutathione
Potassium
Ligands
Poisons
Potassium Channels
NAD
Homeostasis
Escherichia coli

Keywords

  • channel regulation
  • KTN structure
  • Escherichia coli
  • K+ channels
  • RCK domain
  • nucleic acids
  • gating ring
  • activation
  • protein
  • system
  • KEFC
  • transport

Cite this

Roosild, T. P., Castronovo, S., Healy, J., Miller, S., Pliotas, C., Rasmussen, T., ... Booth, I. R. (2010). Mechanism of ligand-gated potassium efflux in bacterial pathogens. PNAS, 107(46), 19784-19789. https://doi.org/10.1073/pnas.1012716107

Mechanism of ligand-gated potassium efflux in bacterial pathogens. / Roosild, Tarmo P (Corresponding Author); Castronovo, Samantha; Healy, Jess; Miller, Samantha; Pliotas, Christos; Rasmussen, Tim; Bartlett, Wendy; Conway, Stuart J; Booth, Ian R (Corresponding Author).

In: PNAS, Vol. 107, No. 46, 16.11.2010, p. 19784-19789.

Research output: Contribution to journalArticle

Roosild, TP, Castronovo, S, Healy, J, Miller, S, Pliotas, C, Rasmussen, T, Bartlett, W, Conway, SJ & Booth, IR 2010, 'Mechanism of ligand-gated potassium efflux in bacterial pathogens', PNAS, vol. 107, no. 46, pp. 19784-19789. https://doi.org/10.1073/pnas.1012716107
Roosild TP, Castronovo S, Healy J, Miller S, Pliotas C, Rasmussen T et al. Mechanism of ligand-gated potassium efflux in bacterial pathogens. PNAS. 2010 Nov 16;107(46):19784-19789. https://doi.org/10.1073/pnas.1012716107
Roosild, Tarmo P ; Castronovo, Samantha ; Healy, Jess ; Miller, Samantha ; Pliotas, Christos ; Rasmussen, Tim ; Bartlett, Wendy ; Conway, Stuart J ; Booth, Ian R. / Mechanism of ligand-gated potassium efflux in bacterial pathogens. In: PNAS. 2010 ; Vol. 107, No. 46. pp. 19784-19789.
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