Evidence for the covalent binding of hydroxyethyl radicals to rat liver microsomal proteins.
It is well known that acetaldehyde is capable of covalent binding to liver proteins. However, in experiments using liver microsomes prepared from chronically ethanol-fed rats we have observed that the addition of EDTA-iron complex to the microsomes increases by about 4-5 fold both the spin trapping of hydroxyethyl radicals and the covalent binding of 14C-ethanol to proteins, while it only doubles acetaldehyde formation. Conversely, the presence of GSH strongly decreases the trapping of hydroxyethyl radicals and completely inhibits the covalent binding, without affecting acetaldehyde production. Furthermore, the spin trapping agent 4-pyridyl-N-oxide-t-butyl nitrone (4-POBN), previously employed for the detection of hydroxyethyl radicals, decreases by about 70% the covalent binding of 14C-ethanol to microsomal proteins. 4-POBN does not affect acetaldehyde production by liver microsomes, nor does it interfere with the covalent binding of acetaldehyde produced by ADH-mediated oxidation of ethanol. The results obtained indicate that hydroxyethyl radicals generated during ethanol oxidation by cytochrome P-450 play an important role in the alkylation of microsomal proteins consequent to ethanol metabolism.
Albano
E
E
Department of Experimental Medicine and Oncology, University of Turin, Torino, Italy.
Parola
M
M
Comoglio
A
A
Dianzani
M U
MU
D000079:Acetaldehyde / Q000493:pharmacokinetics; D000818:Animals; D001665:Binding Sites; D000431:Ethanol / Q000493:pharmacokinetics; D005609:Free Radicals; D017665:Hydroxyl Radical / Q000378:metabolism*; D008108:Liver Diseases, Alcoholic / Q000378:metabolism*; D008297:Male; D008862:Microsomes, Liver / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017382:Reactive Oxygen Species / Q000378:metabolism*
D005609:Free Radicals; D017382:Reactive Oxygen Species; D017665:Hydroxyl Radical; D000431:Ethanol; D000079:Acetaldehyde