Detection of benzoquinone adducts to rat liver protein sulfhydryl groups using specific antibodies

Chemical Research in Toxicology
E M Rombach, R P Hanzlik

Abstract

Benzoquinone is an electrophilic metabolite of bromobenzene and other simple aromatic compounds of toxicological interest including benzene, phenol, hydroquinone, and acetaminophen. In reacting with proteins benzoquinone shows great selectivity for Michael addition to sulfhydryl groups and formation of S-(2,5-dihydroxyphenyl) protein adducts. To facilitate the specific detection and eventual isolation and identification of such adducted proteins, we prepared an antiserum capable of recognizing hydroquinone moieties by immunizing rabbits with keyhole limpet hemocyanin modified with 3-[2,5-dihydroxyphenyl)thio]propanoyl groups as haptens. The antiserum had a high titer and showed high specificity for hapten in competitive ELISA with hapten analogues. In Western blot experiments the antiserum detected not only synthetically haptenized control proteins but also several proteins from rat liver microsomes that had been incubated in vitro with [14C]bromobenzene. This binding was completely blocked by free hapten, showing that it was hapten-specific. Each of the microsomal protein bands detected in the Western blots also contained radioactivity, but not all radioactive protein bands reacted with antibody. This antiserum should prove us...Continue Reading

Citations

Jul 19, 2005·International Immunopharmacology·Nelia D DimitrovaTeodor I Panev
Aug 25, 2005·Journal of Ocular Pharmacology and Therapeutics : the Official Journal of the Association for Ocular Pharmacology and Therapeutics·Steven M Toler
Aug 31, 2000·Biochemical and Biophysical Research Communications·J J Miranda
Aug 5, 2008·Molecular Microbiology·Manuel LiebekeHaike Antelmann
Mar 18, 2004·Bioorganic & Medicinal Chemistry Letters·Weimin YueRobert P Hanzlik
Jun 3, 2006·Journal of Proteome Research·Nigel J WatersJeremy K Nicholson

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