Conjugative metabolism of 1,2-dibromoethane in mitochondria: disruption of oxidative phosphorylation and alkylation of mitochondrial DNA.
1,2-Dibromoethane (DBE) is an environmental contaminant that is metabolized by glutathione S-transferases to a haloethane-glutathione conjugate. Since haloethane-glutathione conjugates are known to alkylate nuclear DNA and cytoplasmic proteins, these effects were investigated in isolated rat liver mitochondria exposed to DBE by measuring guanine adducts and several aspects of oxidative phosphorylation including respiratory control ratios, respiratory enzyme activity, and ATP levels. Mitochondrial large-amplitude swelling and glutathione status were assessed to evaluate mitochondrial membrane integrity and function. When exposed to DBE, mitochondria became uncoupled rapidly, yet no large-amplitude swelling or extramitochondrial glutathione was observed. Mitochondrial GSH was depleted to 2-53% of controls after a 60-min exposure to micromolar quantities of DBE; however, no extramitochondrial GSH or GSSG was detected. The depletion of mitochondrial glutathione corresponded to an increase of an intramitochondrial GSH-conjugate which, based on HPLC elution profiles and retention times, appeared to be S,S'-(1,2-ethanediyl)bis(glutathione). Activities of the NADH oxidase and succinate oxidase respiratory enzyme systems were inhibited 10-74% at micromolar levels of DBE, with succinate oxidase inactivation occurring at lower doses. ATP concentrations in DBE-exposed mitochondria in the presence of succinate were 5-90% lower than in the controls. The DNA adduct S-[2-(N(7)-guanyl)ethyl]glutathione was detected by HPLC in mtDNA isolated from DBE-exposed mitochondria. The results suggest that respiratory enzyme inhibition, glutathione depletion, decreased ATP levels, and DNA alkylation in DBE-exposed mitochondria occur via the formation of an S-(2-bromoethyl)glutathione conjugate, the precursor of the episulfonium ion alkylating species of DBE.
Department of Chemistry, MS-7539, Central Washington University, Ellensburg, WA 98926, USA. cthomas@cwu.edu
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Thomas
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Will
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Schoenberg
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Sanderlin
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Reed
D000255:Adenosine Triphosphate / Q000378:metabolism; D000478:Alkylation / Q000187:drug effects; D000818:Animals; D018736:DNA Adducts / Q000032:analysis; D004272:DNA, Mitochondrial / Q000187:drug effects* / Q000378:metabolism; D004785:Environmental Pollutants / Q000378:metabolism / Q000494:pharmacology; D015946:Ethylene Dibromide / Q000378:metabolism / Q000494:pharmacology*; D005978:Glutathione / Q000172:deficiency / Q000378:metabolism; D006147:Guanine / Q000378:metabolism; D008297:Male; D008930:Mitochondria, Liver / Q000187:drug effects* / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley
D018736:DNA Adducts; D004272:DNA, Mitochondrial; D004785:Environmental Pollutants; D015946:Ethylene Dibromide; D006147:Guanine; D000255:Adenosine Triphosphate; D005978:Glutathione