Metabolism of 3-methylindole in human tissues.
Bioactivation of 3-methylindole (3MI), a highly selective pneumotoxin in goats, was investigated in human lung and liver tissues in order to provide information about the susceptibility of humans to 3MI toxicity. Human lung microsomes were prepared from eight organ transplantation donors and liver microsomes from one of the donors were utilized. The 3MI turnover rate with human lung microsomes was 0.23 +/- 0.06 nmol/mg/min, which was lower than the rate with the human liver microsomes (7.40 nmol/mg/min). The activities were NADPH dependent and inhibited by 1-aminobenzotriazole, a potent cytochrome P-450 suicide substrate inhibitor. Covalent binding of 3MI reactive intermediates to human tissues was determined by incubation of 14C-3MI and NADPH with human lung and liver microsomal proteins. Although human lung microsomes displayed measurable covalent binding activity (2.74 +/- 2.57 pmol/mg/min), the magnitude of this reaction was only 4% as large as that seen with human liver microsomes (62.02 pmol/mg/min). However, the covalent binding was protein dependent and also was inhibited by 1-aminobenzotriazole. Therefore, the bioactivation of 3MI to covalently binding intermediates is catalyzed by cytochrome P-450 in human pulmonary tissues. These activities were compared to those activities measured with tissues from goats. Proteins from goat and human pulmonary and hepatic microsomal incubations were incubated with radioactive 3MI, and radioactive proteins were analyzed by SDS-PAGE and HPLC and visualized by autoradiography and radiochromatography, respectively. The results showed that a 57-kDa protein was clearly the most prominently alkylated target associated with 3MI reactive intermediates.(ABSTRACT TRUNCATED AT 250 WORDS)
Department of Pharmacology and Toxicology, University of Utah, Salt Lake City 84112.
W
W
Ruangyuttikarn
M L
ML
Appleton
G S
GS
Yost
D000328:Adult; D000818:Animals; D001345:Autoradiography; D002675:Child, Preschool; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006041:Goats / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D008168:Lung / Q000378:metabolism*; D008297:Male; D008861:Microsomes / Q000378:metabolism; D008862:Microsomes, Liver / Q000378:metabolism*; D008875:Middle Aged; D011485:Protein Binding; D012862:Skatole / Q000378:metabolism*
D012862:Skatole