Abstract
Acetaminophen is metabolized by cytochrome P450 to a reactive metabolite that covalently binds to proteins and this binding correlates with the hepatotoxicity. The major protein adduct was previously reported to be a 55 kDa protein that was detected on Western blots using antisera specific for 3-(cystein-S-yl)acetaminophen. In this study, the 55 kDa protein was isolated using a combination of ion exchange fast flow chromatography, hydroxyapatite HPLC and anion exchange HPLC. Amino acid sequences of 8 internal peptides from a trypsin digestion of the 55 kDa protein were found to have 97% homology with the deduced amino acid sequence from a cDNA that corresponds to a 56 kDa selenium binding protein. This is the first report of a specific protein to which a metabolite of acetaminophen covalently binds.
MeSH terms
-
Acetaminophen / analogs & derivatives*
-
Acetaminophen / metabolism*
-
Amino Acid Sequence
-
Animals
-
Binding Sites
-
Biotransformation
-
Carrier Proteins / genetics
-
Carrier Proteins / isolation & purification
-
Carrier Proteins / metabolism*
-
Cell Fractionation
-
Chromatography, High Pressure Liquid
-
Chromatography, Ion Exchange
-
Cytosol / metabolism
-
Immune Sera
-
Liver / metabolism*
-
Male
-
Mice
-
Mice, Inbred Strains
-
Molecular Sequence Data
-
Molecular Weight
-
Peptide Fragments / isolation & purification
-
Selenium / isolation & purification
-
Selenium / metabolism*
-
Selenium-Binding Proteins
-
Sequence Homology, Nucleic Acid
-
Ultrafiltration
Substances
-
Carrier Proteins
-
Immune Sera
-
Peptide Fragments
-
Selenbp1 protein, mouse
-
Selenbp2 protein, mouse
-
Selenium-Binding Proteins
-
Acetaminophen
-
3-(cystein-S-yl)paracetamol
-
Selenium