Chromatographic resolution of amino acid adducts of aliphatic halides

R. M. Maiorino, A Jay Gandolfi, K. Brendel, J. R. Mac Donald, I. G. Sipes

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Numerous xenobiotics are known to be bioactivated and to covalently bind to proteins, but the resulting amino acid adducts (AAAs) are unknown. In this study the AAAs of twelve 14C-labeled aliphatic halides were examined after formation in an in vitro microsomal system. After exhaustive solvent extraction of the precipitated microsomal protein, the AAAs were isolated by Pronase digestion, followed by filtration through a 500 mol. wt. exclusion membrane. The liberated AAAs were applied to a constant flow DC-4A cation exchange column, resolved by stepwise buffer elution, collected and counted for radioactivity. Column recovery for applied radioactivity was 100 ± 4%. Generally, 1-4 different AAAs (defined by eluting radioactivity) were resolved, with each organohalogen displaying a characteristic elution profile. Methyl iodide, trichloroethylene and 1,2-dichloroethylene had a single major AAA while bromotrichloromethane, 1,2-dibromoethane, 1,1,1-trichloroethane, 1,2-dichloroethane, 1,1,2-trichloroethane, 2-bromo-2-chloro-1,1,1-trifluoroethane, chloroform and carbon tetrachloride had up to 4 AAAs or more, indicating combinations of binding site(s) and reactive intermediate(s). The single AAA formed following incubation of methyl iodide with the microsomes was identified as S-methylcysteine. Thus, this method appears capable of resolving binding sites and is the initial isolation step for identifying specific adducts to proteins.

Original languageEnglish (US)
Pages (from-to)175-188
Number of pages14
JournalChemico-Biological Interactions
Volume38
Issue number2
DOIs
StatePublished - 1982

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Amino Acids
Radioactivity
Bromotrichloromethane
Ethylene Dibromide
Binding Sites
Trichloroethylene
Pronase
Proteins
Carbon Tetrachloride
Solvent extraction
Xenobiotics
Chloroform
Microsomes
Cations
Digestion
Buffers
Membranes
Recovery
methyl iodide

ASJC Scopus subject areas

  • Toxicology

Cite this

Chromatographic resolution of amino acid adducts of aliphatic halides. / Maiorino, R. M.; Gandolfi, A Jay; Brendel, K.; Mac Donald, J. R.; Sipes, I. G.

In: Chemico-Biological Interactions, Vol. 38, No. 2, 1982, p. 175-188.

Research output: Contribution to journalArticle

Maiorino, R. M. ; Gandolfi, A Jay ; Brendel, K. ; Mac Donald, J. R. ; Sipes, I. G. / Chromatographic resolution of amino acid adducts of aliphatic halides. In: Chemico-Biological Interactions. 1982 ; Vol. 38, No. 2. pp. 175-188.
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