mdr2 encodes P-glycoprotein expressed in the bile canalicular membrane as determined by isoform-specific antibodies

Ellen Buschman, Robert J. Arceci, James Croop, Mingxin Che, Irwin M. Arias, David E. Housman, Philippe Gros

Research output: Contribution to journalArticle

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Abstract

We have produced antibodies specific for the three P-glycoprotein (P-gp) isoforms encoded by the mouse mdr1, mdr2, and mdr3 genes. The anti-Mdr2 and anti-Mdr3 antibodies were generated against synthetic peptides derived from the "linker" region, whereas the anti-Mdr1 antibody was raised against a fusion protein containing the amino terminus of Mdr1. Western blot analysis showed that the three antibodies could discriminate between the three isoforms in membrane fractions from Hamster cells transfected with the corresponding full-length or chimeric mdr cDNAs. Immunocytochemistry studies of mdr-transfected cells showed that the three antibodies specifically recognized each P-gp isoform expressed in whole cells. Immunoblotting of normal mouse tissues revealed that the Mdr2 isoform was expressed at very high levels in liver canalicular membrane vesicles (CMV) but not in membrane vesicles prepared from the basolateral (sinusoidal) domain (SMV). Mdr3 was detected in intestinal brush border membrane vesicles and also in CMV, although at levels much lower than Mdr2. Mdr1 was not detected in CMV or SMV but was detected in endometrial tissue from the gravid uterus. Photolabeling experiments with [125I]iodoarylazidoprazosin followed by immunoprecipitation with isoform-specific antibodies indicated that, in CMV, Mdr3 but not Mdr2 could bind the drug analogue.

Original languageEnglish (US)
Pages (from-to)18093-18099
Number of pages7
JournalJournal of Biological Chemistry
Volume267
Issue number25
StatePublished - Sep 5 1992
Externally publishedYes

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P-Glycoprotein
Bile
Protein Isoforms
Membranes
Antibodies
Anti-Idiotypic Antibodies
Tissue
Brushes
Microvilli
Immunoprecipitation
Immunoblotting
Cricetinae
Liver
Uterus
Fusion reactions
Complementary DNA
Genes
Western Blotting
Immunohistochemistry
Cells

ASJC Scopus subject areas

  • Biochemistry

Cite this

Buschman, E., Arceci, R. J., Croop, J., Che, M., Arias, I. M., Housman, D. E., & Gros, P. (1992). mdr2 encodes P-glycoprotein expressed in the bile canalicular membrane as determined by isoform-specific antibodies. Journal of Biological Chemistry, 267(25), 18093-18099.

mdr2 encodes P-glycoprotein expressed in the bile canalicular membrane as determined by isoform-specific antibodies. / Buschman, Ellen; Arceci, Robert J.; Croop, James; Che, Mingxin; Arias, Irwin M.; Housman, David E.; Gros, Philippe.

In: Journal of Biological Chemistry, Vol. 267, No. 25, 05.09.1992, p. 18093-18099.

Research output: Contribution to journalArticle

Buschman, E, Arceci, RJ, Croop, J, Che, M, Arias, IM, Housman, DE & Gros, P 1992, 'mdr2 encodes P-glycoprotein expressed in the bile canalicular membrane as determined by isoform-specific antibodies', Journal of Biological Chemistry, vol. 267, no. 25, pp. 18093-18099.
Buschman, Ellen ; Arceci, Robert J. ; Croop, James ; Che, Mingxin ; Arias, Irwin M. ; Housman, David E. ; Gros, Philippe. / mdr2 encodes P-glycoprotein expressed in the bile canalicular membrane as determined by isoform-specific antibodies. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 25. pp. 18093-18099.
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