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Absolute immunoquantification of the expression of ABC transporters P-glycoprotein, breast cancer resistance protein and multidrug resistance-associated protein 2 in human liver and duodenum

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Absolute immunoquantification of the expression of ABC transporters P-glycoprotein, breast cancer resistance protein and multidrug resistance-associated protein 2 in human liver and duodenum. / Tucker, Theodora G. H. A.; Milne, Alison M.; Fournel-Gigleux, Sylvie; Fenner, Katherine S.; Coughtrie, Michael W. H.

In: Biochemical Pharmacology, Vol. 83, No. 2, 2012, p. 279-285.

Research output: Contribution to journalArticle

Harvard

Tucker, TGHA, Milne, AM, Fournel-Gigleux, S, Fenner, KS & Coughtrie, MWH 2012, 'Absolute immunoquantification of the expression of ABC transporters P-glycoprotein, breast cancer resistance protein and multidrug resistance-associated protein 2 in human liver and duodenum' Biochemical Pharmacology, vol 83, no. 2, pp. 279-285., 10.1016/j.bcp.2011.10.017

APA

Tucker, T. G. H. A., Milne, A. M., Fournel-Gigleux, S., Fenner, K. S., & Coughtrie, M. W. H. (2012). Absolute immunoquantification of the expression of ABC transporters P-glycoprotein, breast cancer resistance protein and multidrug resistance-associated protein 2 in human liver and duodenum. Biochemical Pharmacology, 83(2), 279-285. 10.1016/j.bcp.2011.10.017

Vancouver

Tucker TGHA, Milne AM, Fournel-Gigleux S, Fenner KS, Coughtrie MWH. Absolute immunoquantification of the expression of ABC transporters P-glycoprotein, breast cancer resistance protein and multidrug resistance-associated protein 2 in human liver and duodenum. Biochemical Pharmacology. 2012;83(2):279-285. Available from: 10.1016/j.bcp.2011.10.017

Author

Tucker, Theodora G. H. A.; Milne, Alison M.; Fournel-Gigleux, Sylvie; Fenner, Katherine S.; Coughtrie, Michael W. H. / Absolute immunoquantification of the expression of ABC transporters P-glycoprotein, breast cancer resistance protein and multidrug resistance-associated protein 2 in human liver and duodenum.

In: Biochemical Pharmacology, Vol. 83, No. 2, 2012, p. 279-285.

Research output: Contribution to journalArticle

Bibtex - Download

@article{85d466c7fef54629840e40bd702b78a4,
title = "Absolute immunoquantification of the expression of ABC transporters P-glycoprotein, breast cancer resistance protein and multidrug resistance-associated protein 2 in human liver and duodenum",
keywords = "Drug transporters, Immunoquantification, BCRP, MRP2, P-glycoprotein, Tandem mass spectrometry, Export pump, Liquid chromatography, Human intestine, Caco-2 cells, Quantification, Gene, Sulfotransferase",
author = "Tucker, {Theodora G. H. A.} and Milne, {Alison M.} and Sylvie Fournel-Gigleux and Fenner, {Katherine S.} and Coughtrie, {Michael W. H.}",
note = "Copyright © 2011 Elsevier Inc. All rights reserved.",
year = "2012",
doi = "10.1016/j.bcp.2011.10.017",
volume = "83",
number = "2",
pages = "279--285",
journal = "Biochemical Pharmacology",
issn = "0006-2952",

}

RIS (suitable for import to EndNote) - Download

TY - JOUR

T1 - Absolute immunoquantification of the expression of ABC transporters P-glycoprotein, breast cancer resistance protein and multidrug resistance-associated protein 2 in human liver and duodenum

A1 - Tucker,Theodora G. H. A.

A1 - Milne,Alison M.

A1 - Fournel-Gigleux,Sylvie

A1 - Fenner,Katherine S.

A1 - Coughtrie,Michael W. H.

AU - Tucker,Theodora G. H. A.

AU - Milne,Alison M.

AU - Fournel-Gigleux,Sylvie

AU - Fenner,Katherine S.

AU - Coughtrie,Michael W. H.

PY - 2012

Y1 - 2012

N2 - <p>The ATP-binding cassette (ABC) transporters breast cancer resistance protein (BCRP), multidrug resistance-associated protein 2 (MRP2), and P-glycoprotein (Pgp) are important in the distribution and elimination of many drugs and endogenous metabolites. Due to their membrane location and hydrophobicity it is difficult to generate purified protein standards to quantify these transporters in human tissues. The present study generated transporter proteins fused with the S-peptide of ribonuclease for use as standards in immunoquantification in human liver and small intestine. Quantification of the S.tag (TM), a 15 amino acid peptide, is based on the formation of a functional ribonuclease activity upon its high affinity reconstitution with ribonuclease S-protein. S-tagged transporters were used as full-length protein standards in the immunoquantification of endogenous BCRP, MRP2, and Pgp levels in 14 duodenum and 13 liver human tissue samples. Expression levels in the duodenum were 305 +/- 248 (BCRP), 66 +/- 70 (MRP2), and 275 +/- 205 (Pgp) fmoles per cm(2). Hepatic levels were 2.6 +/- 0.9 (BCRP), 19.8 +/- 10.5 (MRP2), and 26.1 +/- 10.1 (total Pgp) pmoles per g of liver. The mean hepatic scaling factor was 35.8 mg crude membrane per g of liver, and the mean duodenal scaling factor was 1.3 mg crude membrane per cm(2) mucosal lining. Interindividual variability was greater in duodenal samples than liver samples. It is hoped that this innovative method of quantifying these transporters (and other membrane proteins) will improve in vivo-in vitro extrapolation and in silica prediction of drug absorption and elimination, thus supporting drug development. (C) 2011 Elsevier Inc. All rights reserved.</p>

AB - <p>The ATP-binding cassette (ABC) transporters breast cancer resistance protein (BCRP), multidrug resistance-associated protein 2 (MRP2), and P-glycoprotein (Pgp) are important in the distribution and elimination of many drugs and endogenous metabolites. Due to their membrane location and hydrophobicity it is difficult to generate purified protein standards to quantify these transporters in human tissues. The present study generated transporter proteins fused with the S-peptide of ribonuclease for use as standards in immunoquantification in human liver and small intestine. Quantification of the S.tag (TM), a 15 amino acid peptide, is based on the formation of a functional ribonuclease activity upon its high affinity reconstitution with ribonuclease S-protein. S-tagged transporters were used as full-length protein standards in the immunoquantification of endogenous BCRP, MRP2, and Pgp levels in 14 duodenum and 13 liver human tissue samples. Expression levels in the duodenum were 305 +/- 248 (BCRP), 66 +/- 70 (MRP2), and 275 +/- 205 (Pgp) fmoles per cm(2). Hepatic levels were 2.6 +/- 0.9 (BCRP), 19.8 +/- 10.5 (MRP2), and 26.1 +/- 10.1 (total Pgp) pmoles per g of liver. The mean hepatic scaling factor was 35.8 mg crude membrane per g of liver, and the mean duodenal scaling factor was 1.3 mg crude membrane per cm(2) mucosal lining. Interindividual variability was greater in duodenal samples than liver samples. It is hoped that this innovative method of quantifying these transporters (and other membrane proteins) will improve in vivo-in vitro extrapolation and in silica prediction of drug absorption and elimination, thus supporting drug development. (C) 2011 Elsevier Inc. All rights reserved.</p>

KW - Drug transporters

KW - Immunoquantification

KW - BCRP

KW - MRP2

KW - P-glycoprotein

KW - Tandem mass spectrometry

KW - Export pump

KW - Liquid chromatography

KW - Human intestine

KW - Caco-2 cells

KW - Quantification

KW - Gene

KW - Sulfotransferase

U2 - 10.1016/j.bcp.2011.10.017

DO - 10.1016/j.bcp.2011.10.017

M1 - Article

JO - Biochemical Pharmacology

JF - Biochemical Pharmacology

SN - 0006-2952

IS - 2

VL - 83

SP - 279

EP - 285

ER -

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