title: The critical role of S-lactoylglutathione formation during methylglyoxal detoxification in Escherichia coli.
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 ΔgloB mutant is as tolerant of MG as the parent, despite having the same degree of inhibition of MG detoxification as a ΔgloA 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.
authors.affiliation: School of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK.
authors.forename: Ertan
authors.initials: E
authors.lastname: Ozyamak
authors.forename: Susan S
authors.initials: SS
authors.lastname: Black
authors.forename: Claire A
authors.initials: CA
authors.lastname: Walker
authors.forename: Morag J
authors.initials: MJ
authors.lastname: Maclean
authors.forename: Wendy
authors.initials: W
authors.lastname: Bartlett
authors.forename: Samantha
authors.initials: S
authors.lastname: Miller
authors.forename: Ian R
authors.initials: IR
authors.lastname: Booth
mesh_terms: D004926:Escherichia coli / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism; D005978:Glutathione / Q000031:analogs & derivatives* / Q000378:metabolism; D007791:Lactoylglutathione Lyase / Q000235:genetics / Q000378:metabolism*; D050296:Microbial Viability; D027961:Potassium-Hydrogen Antiporters / Q000235:genetics / Q000378:metabolism; D011765:Pyruvaldehyde / Q000378:metabolism* / Q000494:pharmacology
chemical_list: D029968:Escherichia coli Proteins; D027961:Potassium-Hydrogen Antiporters; C421962:kefB protein, E coli; D011765:Pyruvaldehyde; D007791:Lactoylglutathione Lyase; D005978:Glutathione; C013585:S-lactoylglutathione