Detection of cysteine conjugate metabolite adduct formation with specific mitochondrial proteins using antibodies raised against halothane metabolite adducts.
Antibodies raised against halothane metabolite adducts cross-react with S-(1,1,2,2-tetrafluoroethyl)-L-cysteine (TFEC) and S-(2-chloro-1,1,2-trifluoroethyl)-L-cysteine metabolite adducts. Using these antibodies in immunohistochemical experiments, metabolite binding was localized to the damaged areas of the proximal tubule after treatment of male rats with TFEC. Immunoblot analysis of subcellular fractions of rat kidney tissue after in vivo treatment with TFEC revealed a high specificity for binding of metabolites to proteins of the mitochondrial fraction. These proteins may represent target molecules which play a role in cysteine conjugate induced nephrotoxicity.
W. Alton Jones Cell Science Center, Lake Placid, New York 12946.
P J
PJ
Hayden
T
T
Ichimura
D J
DJ
McCann
L R
LR
Pohl
J L
JL
Stevens
D000081:Acetamides; D000818:Animals; D003429:Cross Reactions; D003545:Cysteine / Q000031:analogs & derivatives* / Q000032:analysis / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D005463:Fluoroacetates; D006221:Halothane / Q000276:immunology*; D006845:Hydrocarbons, Fluorinated / Q000032:analysis* / Q000276:immunology; D015151:Immunoblotting; D007150:Immunohistochemistry; D007674:Kidney Diseases / Q000139:chemically induced / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008928:Mitochondria / Q000378:metabolism*; D051381:Rats; D011919:Rats, Inbred Strains; D014269:Trifluoroacetic Acid / Q000276:immunology; D005466:Fluorocarbons
D000081:Acetamides; D003545:Cysteine; D005463:Fluoroacetates; D006221:Halothane; D006845:Hydrocarbons, Fluorinated; D014269:Trifluoroacetic Acid; C030688:trifluoroacetamide; C064116:S-(1,1,2,2-tetrafluoroethyl)cysteine; D005466:Fluorocarbons; C047601:S-(2-chloro-1,1,2-trifluoroethyl)cysteine