Elsevier

Biochemical Pharmacology

Volume 40, Issue 3, 1 August 1990, Pages 573-579
Biochemical Pharmacology

Immunochemical quantitation of 3-(cystein-S-yl)acetaminophen protein adducts in subcellular liver fractions following a hepatotoxic dose of acetaminophen

https://doi.org/10.1016/0006-2952(90)90558-3Get rights and content

Abstract

The hepatotoxicity of acetaminophen correlates with the formation of 3-(cystein-S-yl)acetaminophen protein adducts. Using a sensitive and specific immunochemical assay, we quantitated the formation of these protein adducts in liver fractions and serum after administration of a hepatotoxic dose of acetaminophen (400 mg/kg) to B6C3F1 mice. Adducts in the cytosolic fraction increased to 3.6 nmol/mg protein at 2 hr and then decreased to 1.1 nmol/mg protein by 8 hr. Concomitant with the decrease in adducts in the cytosol, 3-(cystein-S-yl)acetaminophen protein adducts appeared in serum and their levels paralleled increases in serum alanine aminotransferase. Microsomal protein adducts peaked at 1hr (0.7 nmol/mg protein) and subsequently decreased to 0.2 nmol/mg at 8 hr. The 4000 g pellet (nuclei, plasma membranes, and cell debris) had the highest level of adducts (3.5 nmol/mg protein), which remained constant from 1 to 8 hr. Evaluation of fractions purified from a 960 g pellets indicated that the highest concentration of 3-(cystein-S-yl)acetaminophen protein adducts was located in plasma membranes and mitochondria; peak levels were 10.3 and 5.1 nmol/mg respectively. 3-(Cystein-S-yl) acetaminophen protein adducts were detected in nuclei only after enzymatic hydrolysis of the proteins. The localization of high levels of 3-(cystein-S-yl)acetaminophen protein adducts in plasma membranes and mitochondria may play a critical role in acetaminophen toxicity.

References (29)

  • DGD Davidson et al.

    Acute liver necrosis following overdose of paracetamol

    Br Med J

    (1966)
  • JA Hinson

    Biochemical toxicology of acetaminophen

    Rev Biochem Toxicol

    (1980)
  • JR Mitchell et al.

    Acetaminophen-induced hepatic necrosis. IV. Protective role of glutathione

    J Pharmacol Exp Ther

    (1973)
  • JA Hinson et al.

    3-(Glutathion-S-yl)acetaminophen: A biliary metabolite of acetaminophen

    Drug Metab Dispos

    (1982)
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