Binding of acetaldehyde to a glutathione metabolite: mass spectrometric characterization of an acetaldehyde-cysteinylglycine conjugate.
BACKGROUND
Ethanol administration decreases hepatic glutathione levels and increases urinary sulfhydryl excretion. Ethanol-induced liver injury is blunted by the administration of glutathione precursors. Acetaldehyde generated in the metabolism of ethanol binds to a number of amino acid residues in proteins and peptides, but it does not react readily with glutathione. Due to the possible role of acetaldehyde in cysteine and glutathione homeostasis, we investigated the reaction of acetaldehyde to cysteinylglycine, the dipeptide generated in vivo in the hydrolysis of glutathione by gamma-glutamyltransferase.


METHODS
A conjugate between acetaldehyde and cysteinylglycine was generated under physiologically relevant conditions, both in vitro and in vivo. It was separated by a new reverse-phase high-performance liquid chromatography method and identified by electrospray ionization/ion trap tandem mass spectrometric analysis.


RESULTS
The conjugate with a stoichiometry of 1:1 between cysteinylglycine and acetaldehyde is most rapidly generated in vitro and was identified by mass spectroscopy as 2-methyl-thiazolidine-4-carbonyl-glycine. This thiazolidine derivative is stable in vitro and in biological fluids of rats. The conjugate was present in high concentrations in the bile of rats pretreated with ethanol and an inhibitor of aldehyde dehydrogenase.


CONCLUSIONS
The sequestering of cysteinylglycine by acetaldehyde occurs rapidly under physiologic conditions. Long-lived sulfur-containing biomolecules that incorporate acetaldehyde might affect cysteine and glutathione homeostasis and may also play a protective role by reducing circulating acetaldehyde levels. The acetaldehyde conjugate or its metabolic products could potentially serve as markers of ethanol consumption.
Department of Pathology-Anatomy and Cell Biology and the Alcohol Research Center, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA. eleni.anni@jefferson.edu
Helen
H
Anni
Pavlo
P
Pristatsky
Yedy
Y
Israel
D000079:Acetaldehyde / Q000032:analysis / Q000378:metabolism*; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004151:Dipeptides / Q000032:analysis / Q000378:metabolism*; D005978:Glutathione / Q000032:analysis / Q000378:metabolism*; D008297:Male; D011485:Protein Binding / Q000502:physiology; D051381:Rats; D011917:Rats, Inbred Lew; D017207:Rats, Sprague-Dawley; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D004151:Dipeptides; C028505:cysteinylglycine; D005978:Glutathione; D000079:Acetaldehyde