title: Identification of a rat liver microsomal esterase as a target protein for bromobenzene metabolites.
abstract: The hepatotoxicity of bromobenzene and many other simple organic molecules has been associated with their biotransformation to chemically reactive metabolites and the subsequent covalent binding of those metabolites to cellular macromolecules. To identify proteins targeted by bromobenzene metabolites, we incubated [14C]bromobenzene in vitro with liver microsomes from phenobarbital-induced rats under conditions which typically led to covalent binding of 2-4 nmol equiv of bromobenzene/mg of protein. Microsomal proteins were solubilized with detergent, separated by chromatography and electrophoresis, and analyzed for 14C by phosphorimaging of stained blots. Much of the radioactivity was associated with several bands of proteins of ca. 50-60 kDa, plus another prominent band around 70 kDa, but labeling density appeared to vary considerably overall. A major radiolabeled protein was purified by preparative electrophoresis and submitted to automated Edman microsequencing. Its N-terminal sequence was found to correspond to that of a known rat liver microsomal carboxylesterase (E.C. 3.1.1.1) previously identified as a target for reactive metabolites of halothane. The extent to which covalent modification of this protein by reactive metabolites contributes to the production of hepatotoxic effects remains to be determined.
authors.affiliation: Department of Medicinal Chemistry, University of Kansas, Lawrence 66045-2506, USA.
authors.forename: E M
authors.initials: EM
authors.lastname: Rombach
authors.forename: R P
authors.initials: RP
authors.lastname: Hanzlik
mesh_terms: D000818:Animals; D001969:Bromobenzenes / Q000378:metabolism / Q000633:toxicity*; D002265:Carboxylic Ester Hydrolases / Q000187:drug effects / Q000378:metabolism*; D002852:Chromatography, Ion Exchange; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D001969:Bromobenzenes; D011506:Proteins; C032036:bromobenzene; D002265:Carboxylic Ester Hydrolases