Identification of P-glycoprotein in renal brush border membranes

Debra M. Lieberman; Reinhart A.F. Reithmeier; Victor Ling; Jeffrey H.M. Charuk; Howard Goldberg; Karl L. Skorecki

doi:10.1016/0006-291x(89)91988-8

Identification of P-glycoprotein in renal brush border membranes

Debra M. Lieberman, Reinhart A.F. Reithmeier, Victor Ling, Jeffrey H.M. Charuk, Howard Goldberg, Karl L. Skorecki

Research output: Contribution to journal › Article › peer-review

52 Scopus citations

Abstract

A monoclonal antibody (C219) that recognizes the P-glycoprotein (M_r = 170,000) in plasma membranes of multidrug-resistant Chinese hamster ovary (CHO) cell lines was used to assay renal brush border membrane (BBM) and basolateral membrane (BLM) fractions for the presence of a cross-reactive polypeptide. The C219 antibody bound to a 155,000 dalton protein in immunoblots of rat BBM but not BLM proteins resolved by sodium dodecyl sulfate gel electrophoresis. The corresponding human kidney BBM and dog kidney BBM proteins had molecular weights of 170,000 and 160,000 respectively. The glycoprotein nature of the renal protein was shown by its sensitivity to N-glycanase treatment which reduced the apparent molecular weight of the dog protein to 120,000. In addition, dog P-glycoprotein could be bound to and eluted from immobilized wheat germ agglutinin. The molecular weight, antibody crossreactivity, glycosidase sensitivity and lectin binding show that this protein is a normal kidney analogue of the P-glycoprotein induced in multidrug resistant cell lines.

Original language	English
Pages (from-to)	244-252
Number of pages	9
Journal	Biochemical and Biophysical Research Communications
Volume	162
Issue number	1
DOIs	https://doi.org/10.1016/0006-291x(89)91988-8
State	Published - 14 Jul 1989
Externally published	Yes

Bibliographical note

Funding Information:
ACKNOWLEDGMENTS: This work was funded by grants from the Medical Research Council of Canada to R.A.F.R., V.L. and K.L.S. and a grant from the National cancer Institute to V.L. J.H.M.C. and H.G. are recipients of MRC and Kidney Foundation Postdoctoral Fellowships respectively.

Funding

ACKNOWLEDGMENTS: This work was funded by grants from the Medical Research Council of Canada to R.A.F.R., V.L. and K.L.S. and a grant from the National cancer Institute to V.L. J.H.M.C. and H.G. are recipients of MRC and Kidney Foundation Postdoctoral Fellowships respectively.

Funders	Funder number
National Cancer Institute
Kidney Foundation of Central Pennsylvania
Medical Research Council Canada

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abstract = "A monoclonal antibody (C219) that recognizes the P-glycoprotein (Mr = 170,000) in plasma membranes of multidrug-resistant Chinese hamster ovary (CHO) cell lines was used to assay renal brush border membrane (BBM) and basolateral membrane (BLM) fractions for the presence of a cross-reactive polypeptide. The C219 antibody bound to a 155,000 dalton protein in immunoblots of rat BBM but not BLM proteins resolved by sodium dodecyl sulfate gel electrophoresis. The corresponding human kidney BBM and dog kidney BBM proteins had molecular weights of 170,000 and 160,000 respectively. The glycoprotein nature of the renal protein was shown by its sensitivity to N-glycanase treatment which reduced the apparent molecular weight of the dog protein to 120,000. In addition, dog P-glycoprotein could be bound to and eluted from immobilized wheat germ agglutinin. The molecular weight, antibody crossreactivity, glycosidase sensitivity and lectin binding show that this protein is a normal kidney analogue of the P-glycoprotein induced in multidrug resistant cell lines.",

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AU - Goldberg, Howard

AU - Skorecki, Karl L.

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N2 - A monoclonal antibody (C219) that recognizes the P-glycoprotein (Mr = 170,000) in plasma membranes of multidrug-resistant Chinese hamster ovary (CHO) cell lines was used to assay renal brush border membrane (BBM) and basolateral membrane (BLM) fractions for the presence of a cross-reactive polypeptide. The C219 antibody bound to a 155,000 dalton protein in immunoblots of rat BBM but not BLM proteins resolved by sodium dodecyl sulfate gel electrophoresis. The corresponding human kidney BBM and dog kidney BBM proteins had molecular weights of 170,000 and 160,000 respectively. The glycoprotein nature of the renal protein was shown by its sensitivity to N-glycanase treatment which reduced the apparent molecular weight of the dog protein to 120,000. In addition, dog P-glycoprotein could be bound to and eluted from immobilized wheat germ agglutinin. The molecular weight, antibody crossreactivity, glycosidase sensitivity and lectin binding show that this protein is a normal kidney analogue of the P-glycoprotein induced in multidrug resistant cell lines.

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Identification of P-glycoprotein in renal brush border membranes

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