Acyltransferase-mediated binding of N-hydroxyarylamides to nucleic acids.

N-Hydroxyarylamides are metabolically activated to nucleic acid-binding species by the action of N,O-acyltransferase (AT). The substrate specificity of these enzymes in rat, guinea pig, monkey, baboon, pig, and human liver has been examined by measuring the AT-mediated nucleic acid binding of the N-formyl, N-acetyl, and N-propionyl derivatives of N-hydroxy-2-aminofluorene. Human and pig enzymes catalyzed binding in the order formyl greater than acetyl greater than propionyl, while for the other species the order was acetyl greater than propionyl greater than formyl. Ammonium sulfate fractionation of the cytosols suggested that the baboon and rat have at least two different AT's: one with a higher specificity for the formyl derivative; the other with a marked preference for acetyl and propionyl compounds. Only one form, with a high formyl group specificity, was detected from human liver. The identity of the in vitro AT-mediated DNA adducts from rat, baboon, and human liver was established. In each instance, one adduct accounted for greater than 75% of the bound 2-aminofluorene (AF) residues. This product had a high-pressure liquid chromatography retention time and pH-dependent partition characteristics identical to those of an adduct synthesized by an acid-dependent (pH 4.6) reaction of N-hydroxy-2-aminofluorene with calf thymus DNA. This synthetic adduct has been identified as N-(deoxyguanosin-8-yl)-2-aminofluorene by nuclear magnetic resonance, mass, and ultraviolet light spectroscopy. Moreover, it was identical to the product obtained from the alkaline (pH 12) hydrolysis of N-(deoxyguanosin-8-yl)-2-acetylaminofluorene. Since an arylaminated (i.e., aminofluorene) residue(s) is the major product found in rat liver DNA following administration of N-hydroxy-N-acetyl-2-aminofluorene, these data suggest that AT may play a major role in the formation of this DNA-carcinogen adduct.