Evidence for the formation of Michael adducts from reactions of (E,E)-muconaldehyde with glutathione and other thiols

Department of Environmental Sciences and Engineering, University of North Carolina at Chapel Hill, North Carolina, United States
Bioorganic Chemistry (Impact Factor: 2.15). 11/2005; 33(5):363-73. DOI: 10.1016/j.bioorg.2005.05.004
Source: PubMed


Glutathione induces the rapid isomerization of (Z,Z)-muconaldehyde to (E,E)-muconaldehyde via (E,Z)-muconaldehyde, probably via reversible Michael addition of the thiol to one of the enal moieties of the muconaldehyde. Reactions of (E,E)-muconaldehyde with glutathione (in the presence and absence of equine glutathione S-transferase), phenylmethanethiol, N-acetyl-l-cysteine, and N-acetyl-l-cysteine methyl ester were investigated using mass spectrometric techniques. In each case, evidence was obtained for the formation of Michael adducts, e.g., reaction between (E,E)-muconaldehyde and glutathione gave 4-glutathionyl-hex-2-enedial and 3,4-bis-glutathionyl-hexanedial. These experiments suggest that (Z,Z)-muconaldehyde, a putative metabolite of benzene, could lead to the long established urinary metabolite of benzene, (E,E)-muconic acid, via glutathione-mediated isomerization to (E,E)-muconaldehyde.

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    • "Human exposures to benzene at air concentrations between 0.1 and 10 ppm, result in urinary metabolite profiles with 70–85% PH, 5–10% each of HQ, MA and CAT, and less than 1% of S-PMA [8]. Benzene oxide, the benzoquinones , muconaldehydes, and benzene diol epoxides (formed from CYP oxidation of benzene dihydrodiol) are electrophiles that readily react with peptides and proteins [9] [10] [11] [12] and can thereby interfere with cellular function [13]. The identification of metabolic susceptibility factors has confirmed the importance of metabolism in benzene toxicity. "
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    ABSTRACT: Benzene exposure is associated with acute myeloid leukemia (AML), myelodysplastic syndromes (MDS), and probably lymphoma and childhood leukemia. Biological plausibility for a causal role of benzene in these diseases comes from its toxicity to hematopoietic stem cells (HSC) or progenitor cells, from which all leukemias and related disorders arise. The effect of this toxicity is manifest as lowered blood counts (hematotoxicity), even in individuals occupationally exposed to low levels of benzene. Benzene can induce AML/MDS via several well-characterized pathways associated with these diseases. Through its metabolites, benzene induces multiple alterations that likely contribute to the leukemogenic process, and appears to operate via multiple modes of action. To improve mechanistic understanding and for risk assessment purposes, it may be possible to measure several of the key events in these modes of action in an in vitro model of the bone marrow stem cell niche. Even though benzene is leukemogenic at relatively low occupational levels of exposure, it seems unlikely that it is a major cause of leukemia in the general population exposed to benzene in the ppb range. Other established non-genetic causes of AML, e.g. smoking, ionizing radiation and cancer chemotherapy, also only explain about 20% of AML incidence, leaving ∼80% unexplained. The question arises as to how to find the causes of the majority of de novo AMLs that remain unexplained. We propose that we should attempt to characterize the 'exposome' of human leukemia by using unbiased laboratory-based methods to find the unknown 'environmental' factors that contribute to leukemia etiology.
    Full-text · Article · Feb 2011 · Chemico-biological interactions
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    • "Commercially available reduced glutathione (GSH) represents a very convenient starting material for the synthesis of conjugates. Michael addition of glutathione on conjugate aldehyde (Henderson et al., 2005;Schmidt, Ak, & Mrowietz, 2007) and on o-quinones (Modica, Zanaletti, Freccero, & Mella, 2001;Nishizawa et al., 2006) is well-documented. Consequently, we envisaged using directly glutathione as the substrate for the nucleophilic addition of free thiol on trans-2-hexenal in water with a slightly alkaline pH to favour the formation of thiolate ion (data not shown). "
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    ABSTRACT: The isotopically labelled S-3-(hexan-1-ol)-glutathione d2/d3 (G3MHd2/d3) was synthesized with a strategy based on acid-labile protecting groups. The natural analogue of this compound could be a precursor of 3-mercaptohexanol, a varietal thiol reminiscent of grape fruit, released during alcoholic fermentation. In a first time, deuterated glutathione conjugate was used to perform identification and quantification by stable isotope dilution assay of G3MH in musts from several varieties: Sauvignon, Riesling and Gewurztraminer.In a second time, synthetic and natural musts (Sauvignon Blanc) were spiked by G3MHd2/d3 at different levels from 12 to 150 μg L−1 to demonstrate the relationship between 3-mercaptohexan-1-ol (3MH) and G3MH. GC–ITMS/MS analysis of the resulting wines clearly showed the formation of the corresponding deuterated 3MH, that proves the direct connection with G3MH under enological conditions. In Sauvignon Blanc grape juice, a conversion yield of 4.4% was found in duplicate for an initial spiking of G3MH d2/d3 equivalent to 12 μg L−1.
    Full-text · Article · Aug 2010 · Food Chemistry
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    • "air concentrations between 0.1 and 10 ppm, phenol represents 70–85% of urinary benzene metabolites, whereas hydroquinone, muconic acid, and catechol each represent 5–10%, and SPMA represents less than 1% (Kim et al. 2006b). Benzene oxide, the benzoquinones, muconaldehydes, and benzene diol epoxides (formed from CYP oxidation of benzene dihydrodiol ) are electrophiles that readily react with peptides and proteins (Bechtold et al. 1992; Henderson et al. 2005; McDonald et al. 1993; Waidyanatha et al. 2005) and can thereby interfere with cellular function (Smith 1996). Reactions between 1,4-benzoquinone and critical nucleophilic loci in topoisomerase II, an important enzyme involved in DNA replication and maintenance, are mentioned as likely key events in the cascade of effects related to benzene's ability to damage chromosomes (Chen and Eastmond 1995; Lindsey et al. 2004). "
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    ABSTRACT: Recent evidence has shown that humans metabolize benzene more efficiently at environmental air concentrations than at concentrations > 1 ppm. This led us to speculate that an unidentified metabolic pathway was mainly responsible for benzene metabolism at ambient levels. We statistically tested whether human metabolism of benzene is better fitted by a kinetic model having two pathways rather than one. We fit Michaelis-Menten-like models to levels of urinary benzene metabolites and the corresponding air concentrations for 263 nonsmoking Chinese females. Estimated benzene concentrations ranged from less than 0.001 ppm to 299 ppm, with 10th and 90th percentile values of 0.002 ppm and 8.97 ppm, respectively. Using values of Akaike's information criterion obtained under the two models, we found strong statistical evidence favoring two metabolic pathways, with respective affinities (benzene air concentrations analogous to K(m) values) of 301 ppm for the low-affinity pathway (probably dominated by cytochrome P450 enzyme 2E1) and 0.594 ppm for the high-affinity pathway (unknown). The exposure-specific metabolite level predicted by our two-pathway model at nonsaturating concentrations was 184 muM/ppm of benzene, a value close to an independent estimate of 194 muM/ppm for a typical nonsmoking Chinese female. Our results indicate that a nonsmoking woman would metabolize about three times more benzene from the ambient environment under the two-pathway model (184 muM/ppm) than under the one-pathway model (68.6 muM/ppm). In fact, 73% of the ambient benzene dose would be metabolized via the unidentified high-affinity pathway. Because regulatory risk assessments have assumed nonsaturating metabolism of benzene in persons exposed to air concentrations well above 10 ppm, our findings suggest that the true leukemia risks could be substantially greater than currently thought at ambient levels of exposure-about 3-fold higher among nonsmoking females in the general population.
    Full-text · Article · Jun 2009 · Environmental Health Perspectives
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