The calcium-binding protein calreticulin is covalently modified in rat liver by a reactive metabolite of the inhalation anesthetic halothane.
A general procedure is presented for the isolation of several liver microsomal target proteins of the reactive trifluoroacetyl halide metabolite of halothane. It was found that most of these proteins could be selectively extracted from microsomes with 0.1% sodium deoxycholate and separated into partially purified fractions by DEAE-Sepharose anion-exchange chromatography. Using this method, we describe the isolation and identification of a 63-kDa target protein of halothane in rat liver. Amino acid sequences of the N-terminal and of several internal peptides of the protein, as well as the deduced amino acid sequence of a nearly full-length rat liver cDNA clone of the protein, showed 98% identity with a reported murine cDNA that encodes for calreticulin, a major calcium-binding protein of the lumen of endoplasmic reticulum. Although it remains to be determined what role calreticulin has in the development of halothane hepatitis, this study has shown that calreticulin can be a target of reactive metabolites of xenobiotics.
Butler
L E
LE
Laboratory of Chemical Pharmacology, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892.
Thomassen
D
D
Martin
J L
JL
Martin
B M
BM
Kenna
J G
JG
Pohl
L R
LR
D000595:Amino Acid Sequence; D000818:Animals; D002135:Calcium-Binding Proteins / Q000302:isolation & purification / Q000378:metabolism*; D037282:Calreticulin; D002851:Chromatography, High Pressure Liquid; D005463:Fluoroacetates; D006221:Halothane / Q000378:metabolism / Q000494:pharmacology*; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000378:metabolism; D008969:Molecular Sequence Data; D051381:Rats; D011919:Rats, Inbred Strains; D014269:Trifluoroacetic Acid / Q000378:metabolism
D002135:Calcium-Binding Proteins; D037282:Calreticulin; D005463:Fluoroacetates; D014269:Trifluoroacetic Acid; D006221:Halothane