UDP-glucose:glycoprotein glucosyltransferase associates with endoplasmic reticulum chaperones and its activity is decreased in vivo by the inhalation anesthetic halothane.
Halothane causes an idiosyncratic hepatitis that is thought to result, in part, from immune reactions against one or more lumenal endoplasmic reticulum (ER) proteins that have been covalently modified by the trifluoroacetyl chloride metabolite of halothane. In this study, we have identified a 170 kDa protein target of halothane in the liver of rats. The 170 kDa protein was first detected when proteins in lysates of hepatocytes from halothane-treated rats were immunoprecipitated with antisera against several resident ER proteins. This 170 kDa protein was found to be associated with other protein targets of halothane, including protein disulfide isomerase, a protein disulfide isomerase isoform, a 59 kDa carboxylesterase, and 78 kDa glucose-regulated protein. Immunoblotting with antiserum directed against the trifluoroacetylated hapten indicated that the 170 kDa protein was trifluoroacetylated. Based upon its subcellular localization, molecular mass, N-terminal amino acid sequence, and antigenicity, the trifluoroacetylated 170 kDa protein was identified as UDP-glucose:glycoprotein glucosyltransferase (UGGT), a lumenal ER protein that is thought to have a role in the folding of N-linked glycoproteins. Moreover, treatment of rats with halothane caused a 44% decrease in the activity of liver microsomal UGGT, and at least 36% of the change in the activity of the enzyme could be due to a decrease in the level of the protein. The results suggest that the function of UGGT in folding of N-linked glycoproteins may be affected by other resident ER proteins or xenobiotics such as halothane.
Amouzadeh
H R
HR
Molecular and Cellular Toxicology Section, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, Maryland 20892-1760, USA. Hamid_Amouzadeh@nih.gov
Bourdi
M
M
Martin
J L
JL
Martin
B M
BM
Pohl
L R
LR
D018685:Anesthetics, Inhalation / Q000494:pharmacology*; D000818:Animals; D004721:Endoplasmic Reticulum / Q000201:enzymology*; D005964:Glucosyltransferases / Q000032:analysis / Q000037:antagonists & inhibitors* / Q000737:chemistry; D006221:Halothane / Q000494:pharmacology*; D015151:Immunoblotting; D008099:Liver / Q000187:drug effects / Q000201:enzymology; D008297:Male; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology; D018832:Molecular Chaperones / Q000378:metabolism*; D011233:Precipitin Tests; D051381:Rats; D017207:Rats, Sprague-Dawley; D014269:Trifluoroacetic Acid
D018685:Anesthetics, Inhalation; D018832:Molecular Chaperones; D014269:Trifluoroacetic Acid; D005964:Glucosyltransferases; C072423:mannosylglycoprotein 1,3-glucosyltransferase; D006221:Halothane