The topography of trifluoroacetylated protein antigens in liver microsomal fractions from halothane treated rats

J. Gerald Kenna, Jackie L. Martin, Lance R. Pohl

Research output: Contribution to journalArticle

Abstract

Sera from patients with halothane hepatitis contain immunoglobulin G (IgG) antibodies to trifluoroacetylated liver microsomal proteins of 100, 76, 59, 57 and 54 kDa, which are produced as a consequence of metabolism of halothane to trifluoroacetyl halide by cytochrome(s) P450. In the present study, the membrane topographies of the various antigens in rat liver microsomal fractions were investigated. Liver microsomal fractions from rats treated with halothane in vivo, and rat liver microsomal fractions which had been incubated with halothane in vitro, were used as the source of tri-fluoroacetyl antigens. The antigens were detected by immunoblotting. Whereas the 100, 76, 59 and 57 kDa antigens were solubilized from the microsomal membrane by either 0.1 M sodium carbonate or 0.1% (w/v) sodium deoxycholate, the 54 kDa antigen was not solubilized by 0.1% (w/v) sodium deoxycholate. In intact microsomal fractions, the 100, 76, 59 and 57 kDa antigens were not degraded appreciably by trypsin unless detergent was added to permeabilize the microsomal membrane. These results indicate that the 54 kDa antigen is an integral membrane protein, whereas the 100, 76, 59 and 57 kDa antigens are peripheral membrane proteins situated within the lumen of microsomal vesicles, and hence presumably located within the lumen of the endoplasmic reticulum in vivo.

Original languageEnglish (US)
Pages (from-to)621-629
Number of pages9
JournalBiochemical Pharmacology
Volume44
Issue number4
DOIs
StatePublished - Aug 18 1992

Fingerprint

Halothane
Liver
Topography
Rats
Antigens
Proteins
Deoxycholic Acid
Membranes
Membrane Proteins
Immunoblotting
Metabolism
Endoplasmic Reticulum
Detergents
Trypsin
Cytochrome P-450 Enzyme System
Immunoglobulin G
Antibodies
Serum

ASJC Scopus subject areas

  • Pharmacology

Cite this

The topography of trifluoroacetylated protein antigens in liver microsomal fractions from halothane treated rats. / Kenna, J. Gerald; Martin, Jackie L.; Pohl, Lance R.

In: Biochemical Pharmacology, Vol. 44, No. 4, 18.08.1992, p. 621-629.

Research output: Contribution to journalArticle

Kenna, J. Gerald ; Martin, Jackie L. ; Pohl, Lance R. / The topography of trifluoroacetylated protein antigens in liver microsomal fractions from halothane treated rats. In: Biochemical Pharmacology. 1992 ; Vol. 44, No. 4. pp. 621-629.
@article{995612ee802c41d39ba61be1fbbc9561,
title = "The topography of trifluoroacetylated protein antigens in liver microsomal fractions from halothane treated rats",
abstract = "Sera from patients with halothane hepatitis contain immunoglobulin G (IgG) antibodies to trifluoroacetylated liver microsomal proteins of 100, 76, 59, 57 and 54 kDa, which are produced as a consequence of metabolism of halothane to trifluoroacetyl halide by cytochrome(s) P450. In the present study, the membrane topographies of the various antigens in rat liver microsomal fractions were investigated. Liver microsomal fractions from rats treated with halothane in vivo, and rat liver microsomal fractions which had been incubated with halothane in vitro, were used as the source of tri-fluoroacetyl antigens. The antigens were detected by immunoblotting. Whereas the 100, 76, 59 and 57 kDa antigens were solubilized from the microsomal membrane by either 0.1 M sodium carbonate or 0.1{\%} (w/v) sodium deoxycholate, the 54 kDa antigen was not solubilized by 0.1{\%} (w/v) sodium deoxycholate. In intact microsomal fractions, the 100, 76, 59 and 57 kDa antigens were not degraded appreciably by trypsin unless detergent was added to permeabilize the microsomal membrane. These results indicate that the 54 kDa antigen is an integral membrane protein, whereas the 100, 76, 59 and 57 kDa antigens are peripheral membrane proteins situated within the lumen of microsomal vesicles, and hence presumably located within the lumen of the endoplasmic reticulum in vivo.",
author = "Kenna, {J. Gerald} and Martin, {Jackie L.} and Pohl, {Lance R.}",
year = "1992",
month = "8",
day = "18",
doi = "10.1016/0006-2952(92)90395-Y",
language = "English (US)",
volume = "44",
pages = "621--629",
journal = "Biochemical Pharmacology",
issn = "0006-2952",
publisher = "Elsevier Inc.",
number = "4",

}

TY - JOUR

T1 - The topography of trifluoroacetylated protein antigens in liver microsomal fractions from halothane treated rats

AU - Kenna, J. Gerald

AU - Martin, Jackie L.

AU - Pohl, Lance R.

PY - 1992/8/18

Y1 - 1992/8/18

N2 - Sera from patients with halothane hepatitis contain immunoglobulin G (IgG) antibodies to trifluoroacetylated liver microsomal proteins of 100, 76, 59, 57 and 54 kDa, which are produced as a consequence of metabolism of halothane to trifluoroacetyl halide by cytochrome(s) P450. In the present study, the membrane topographies of the various antigens in rat liver microsomal fractions were investigated. Liver microsomal fractions from rats treated with halothane in vivo, and rat liver microsomal fractions which had been incubated with halothane in vitro, were used as the source of tri-fluoroacetyl antigens. The antigens were detected by immunoblotting. Whereas the 100, 76, 59 and 57 kDa antigens were solubilized from the microsomal membrane by either 0.1 M sodium carbonate or 0.1% (w/v) sodium deoxycholate, the 54 kDa antigen was not solubilized by 0.1% (w/v) sodium deoxycholate. In intact microsomal fractions, the 100, 76, 59 and 57 kDa antigens were not degraded appreciably by trypsin unless detergent was added to permeabilize the microsomal membrane. These results indicate that the 54 kDa antigen is an integral membrane protein, whereas the 100, 76, 59 and 57 kDa antigens are peripheral membrane proteins situated within the lumen of microsomal vesicles, and hence presumably located within the lumen of the endoplasmic reticulum in vivo.

AB - Sera from patients with halothane hepatitis contain immunoglobulin G (IgG) antibodies to trifluoroacetylated liver microsomal proteins of 100, 76, 59, 57 and 54 kDa, which are produced as a consequence of metabolism of halothane to trifluoroacetyl halide by cytochrome(s) P450. In the present study, the membrane topographies of the various antigens in rat liver microsomal fractions were investigated. Liver microsomal fractions from rats treated with halothane in vivo, and rat liver microsomal fractions which had been incubated with halothane in vitro, were used as the source of tri-fluoroacetyl antigens. The antigens were detected by immunoblotting. Whereas the 100, 76, 59 and 57 kDa antigens were solubilized from the microsomal membrane by either 0.1 M sodium carbonate or 0.1% (w/v) sodium deoxycholate, the 54 kDa antigen was not solubilized by 0.1% (w/v) sodium deoxycholate. In intact microsomal fractions, the 100, 76, 59 and 57 kDa antigens were not degraded appreciably by trypsin unless detergent was added to permeabilize the microsomal membrane. These results indicate that the 54 kDa antigen is an integral membrane protein, whereas the 100, 76, 59 and 57 kDa antigens are peripheral membrane proteins situated within the lumen of microsomal vesicles, and hence presumably located within the lumen of the endoplasmic reticulum in vivo.

UR - http://www.scopus.com/inward/record.url?scp=0026640574&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026640574&partnerID=8YFLogxK

U2 - 10.1016/0006-2952(92)90395-Y

DO - 10.1016/0006-2952(92)90395-Y

M3 - Article

C2 - 1510711

AN - SCOPUS:0026640574

VL - 44

SP - 621

EP - 629

JO - Biochemical Pharmacology

JF - Biochemical Pharmacology

SN - 0006-2952

IS - 4

ER -