The critical role of S-lactoylglutathione formation during methylglyoxal detoxification in Escherichia coli

Ertan Ozyamak, Susan S Black, Claire A Walker, Morag J Maclean, Wendy Bartlett, Samantha Miller, Ian R Booth

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Survival of exposure to methylglyoxal (MG) in Gram-negative pathogens is largely dependent upon the operation of the glutathione-dependent glyoxalase system, consisting of two enzymes, GlxI (gloA) and GlxII (gloB). In addition, the activation of the KefGB potassium efflux system is maintained closed by glutathione (GSH) and is activated by S-lactoylGSH (SLG), the intermediate formed by GlxI and destroyed by GlxII. Escherichia coli mutants lacking GlxI are known to be extremely sensitive to MG. In this study we demonstrate that a ¿gloB mutant is as tolerant of MG as the parent, despite having the same degree of inhibition of MG detoxification as a ¿gloA strain. Increased expression of GlxII from a multicopy plasmid sensitizes E. coli to MG. Measurement of SLG pools, KefGB activity and cytoplasmic pH shows these parameters to be linked and to be very sensitive to changes in the activity of GlxI and GlxII. The SLG pool determines the activity of KefGB and the degree of acidification of the cytoplasm, which is a major determinant of the sensitivity to electrophiles. The data are discussed in terms of how cell fate is determined by the relative abundance of the enzymes and KefGB.
Original languageEnglish
Pages (from-to)1577-1590
Number of pages14
JournalMolecular Microbiology
Volume78
Issue number6
Early online date29 Oct 2010
DOIs
Publication statusPublished - Dec 2010

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Pyruvaldehyde
Escherichia coli
Glutathione
Enzymes
Potassium
Cytoplasm
Plasmids
S-lactoylglutathione

Keywords

  • Escherichia coli
  • Escherichia coli proteins
  • glutathione
  • lactoylglutathione lyase
  • microbial viability
  • potassium-hydrogen antiporters
  • pyruvaldehyde

Cite this

The critical role of S-lactoylglutathione formation during methylglyoxal detoxification in Escherichia coli. / Ozyamak, Ertan; Black, Susan S; Walker, Claire A; Maclean, Morag J; Bartlett, Wendy; Miller, Samantha; Booth, Ian R.

In: Molecular Microbiology, Vol. 78, No. 6, 12.2010, p. 1577-1590.

Research output: Contribution to journalArticle

Ozyamak, Ertan ; Black, Susan S ; Walker, Claire A ; Maclean, Morag J ; Bartlett, Wendy ; Miller, Samantha ; Booth, Ian R. / The critical role of S-lactoylglutathione formation during methylglyoxal detoxification in Escherichia coli. In: Molecular Microbiology. 2010 ; Vol. 78, No. 6. pp. 1577-1590.
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AB - Survival of exposure to methylglyoxal (MG) in Gram-negative pathogens is largely dependent upon the operation of the glutathione-dependent glyoxalase system, consisting of two enzymes, GlxI (gloA) and GlxII (gloB). In addition, the activation of the KefGB potassium efflux system is maintained closed by glutathione (GSH) and is activated by S-lactoylGSH (SLG), the intermediate formed by GlxI and destroyed by GlxII. Escherichia coli mutants lacking GlxI are known to be extremely sensitive to MG. In this study we demonstrate that a ¿gloB mutant is as tolerant of MG as the parent, despite having the same degree of inhibition of MG detoxification as a ¿gloA strain. Increased expression of GlxII from a multicopy plasmid sensitizes E. coli to MG. Measurement of SLG pools, KefGB activity and cytoplasmic pH shows these parameters to be linked and to be very sensitive to changes in the activity of GlxI and GlxII. The SLG pool determines the activity of KefGB and the degree of acidification of the cytoplasm, which is a major determinant of the sensitivity to electrophiles. The data are discussed in terms of how cell fate is determined by the relative abundance of the enzymes and KefGB.

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