Studies on Quinone-Thioethers. I. Mechanism of Formation and Properties of Thiodione<sup>*</sup>

Nickerson, Walter J.; Falcone, Giuseppe; Strauss, George

doi:10.1021/bi00903a025

Cited by 132 publications

(54 citation statements)

References 11 publications

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“…The formation of a GSH-menadione conjugate has been observed by others (Nickerson et al, 1963). However, we saw no resonances that could be ascribed to such a conjugate in the NMR spectra of CHO whole cells.…”

Section: Resultssupporting

confidence: 62%

“…However, we saw no resonances that could be ascribed to such a conjugate in the NMR spectra of CHO whole cells. The formation of the menadione-GSH conjugate is sensitive to the relative concentrations of menadione and glutathione (Nickerson et al, 1963). It would appear that in the model used here the conditions were not conducive to the formation of a conjugate.…”

Section: Resultsmentioning

confidence: 89%

“…Glutathione reductase catalyses the regeneration of GSH, which maintains the cell in a reduced state. Menadione also interacts with glutathione to form a menadione-GSH conjugate (Nickerson et al, 1963) and with protein-SH groups to form arylated protein (Di Monte et al, 1984).…”

mentioning

confidence: 99%

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Menadione-resistant Chinese hamster ovary cells have an increased capacity for glutathione synthesis

Vallis

Reglinski²,

Garner³

et al. 1997

Br J Cancer

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Summary A cell line (MRc4O) resistant to the model quinone compound, menadione, has been isolated from a parental Chinese hamster ovary cell line (CHO-K1). The known relationship between menadione toxicity and glutathione (GSH) depletion led us to investigate whether the mechanism of resistance of MRc40 was related to alteration in GSH homeostasis. Intracellular concentrations of GSH and cysteine (CySH) were twofold and 3.2-fold greater in MRc40 than in CHO-Kl. Following exposure to menadione, GSH and CySH were depleted, but subsequent recovery of thiols was more rapid and of greater magnitude in MRc40 than in CHO-Kl. Twelve hours after exposure to menadione, the concentrations of GSH and CySH were 9.7-and 4.2-fold greater in MRc40 than in CHO-Kl. Using nuclear magnetic resonance (NMR) spectroscopy, we observed the in situ removal of menadione from cell suspensions of CHO-Kl and MRc40. However, only in CHO-Kl did we observe concomitant depletion of NMR-visible GSH. We conclude that the perturbation of GSH metabolism contributes to the resistant phenotype and is an important characteristic of menadione-resistant CHO cells.

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Section: Resultssupporting

confidence: 62%

Section: Resultsmentioning

confidence: 89%

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Menadione-resistant Chinese hamster ovary cells have an increased capacity for glutathione synthesis

Vallis

Reglinski²,

Garner³

et al. 1997

Br J Cancer

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“…Specificity of wild-type and Cys227-substituted carbonyl reductases for menadione, PGA 1 and the corresponding GSH adducts. GSH adducts were synthesized according to [2] and [17]. Values are the mean of 2±3 determinations, and standard deviations were , 20%.…”

Section: Discussionmentioning

confidence: 99%

“…The GSH adducts of PGA 1 (PGA 1 -SG) and menadione (menadione-SG) were prepared as described by Wermuth [2] and Nickerson et al [17], respectively. Daunorubicin was kindly provided by Rho Ãne-Poulenc.…”

Section: Experimental Procedures Materialsmentioning

confidence: 99%

Identification of the reactive cysteine residue (Cys227) in human carbonyl reductase

Tinguely

Wermuth

1999

European Journal of Biochemistry

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Carbonyl reductase is highly susceptible to inactivation by organomercurials suggesting the presence of a reactive cysteine residue in, or close to, the active site. This residue is also close to a site which binds glutathione. Structurally, carbonyl reductase belongs to the short-chain dehydrogenase/reductase family and contains five cysteine residues, none of which is conserved within the family. In order to identify the reactive residue and investigate its possible role in glutathione binding, alanine was substituted for each cysteine residue of human carbonyl reductase by site-directed mutagenesis. The mutant enzymes were expressed in Escherichia coli and purified to homogeneity. Four of the five mutants (C26A, C122A C150A and C226A) exhibited wild-type-like enzyme activity, although K m values of C226A for three structurally different substrates were increased threefold to 10-fold. The fifth mutant, C227A, showed a 10±15-fold decrease in k cat and a threefold to 40-fold increase in K m , resulting in a 30±500-fold drop in k cat /K m . NaCl (300 mm) increased the activity of C227A 16-fold, whereas the activity of the wild-type enzyme was only doubled. Substitution of serine rather than alanine for Cys227 similarly affected the kinetic constants with the exception that NaCl did not activate the enzyme. Both C227A and C227S mutants were insensitive to inactivation by 4-hydroxymercuribenzoate. Unlike the parent carbonyl compounds, the glutathione adducts of menadione and prostaglandin A 1 were better substrates for the C227A and C227S mutants than the wild-type enzyme. Conversely, the binding of free glutathione to both mutants was reduced. Our findings indicate that Cys227 is the reactive residue and suggest that it is involved in the binding of both substrate and glutathione.

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Trametes versicolor glutathione transferase Xi 3, a dual Cys‐GST with catalytic specificities of both Xi and Omega classes

et al. 2018

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Glutathione transferases (GSTs) from the Xi and Omega classes have a catalytic cysteine residue, which gives them reductase activities. Until now, they have been assigned distinct substrates. While Xi GSTs specifically reduce glutathionyl-(hydro)quinones, Omega GSTs are specialized in the reduction of glutathionyl-acetophenones. Here, we present the biochemical and structural analysis of TvGSTX1 and TvGSTX3 isoforms from the wood-degrading fungus Trametes versicolor. TvGSTX1 reduces GS-menadione as expected, while TvGSTX3 reduces both Xi and Omega substrates. An in-depth structural analysis indicates a broader active site for TvGSTX3 due to specific differences in the nature of the residues situated in the C-terminal helix α9. This feature could explain the catalytic duality of TvGSTX3. Based on phylogenetic analysis, we propose that this duality might exist in saprophytic fungi and ascomycetes.

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Studies on Quinone-Thioethers. I. Mechanism of Formation and Properties of Thiodione^*

Cited by 132 publications

References 11 publications

Menadione-resistant Chinese hamster ovary cells have an increased capacity for glutathione synthesis

Menadione-resistant Chinese hamster ovary cells have an increased capacity for glutathione synthesis

Identification of the reactive cysteine residue (Cys227) in human carbonyl reductase

Trametes versicolor glutathione transferase Xi 3, a dual Cys‐GST with catalytic specificities of both Xi and Omega classes

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Studies on Quinone-Thioethers. I. Mechanism of Formation and Properties of Thiodione*

Cited by 132 publications

References 11 publications

Menadione-resistant Chinese hamster ovary cells have an increased capacity for glutathione synthesis

Menadione-resistant Chinese hamster ovary cells have an increased capacity for glutathione synthesis

Identification of the reactive cysteine residue (Cys227) in human carbonyl reductase

Trametes versicolor glutathione transferase Xi 3, a dual Cys‐GST with catalytic specificities of both Xi and Omega classes

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Studies on Quinone-Thioethers. I. Mechanism of Formation and Properties of Thiodione^*