Transport of Leukotriene C4 by a Cysteine-less Multidrug Resistance Protein 1 (MRP1)

Overview

Journal Biochem J

Specialty Biochemistry

Date 2002 Oct 26

PMID 12398766

Citations 3

Authors

Sung H Lee

Guillermo A Altenberg

Affiliations

Soon will be listed here.

Abstract

Overexpression of multidrug resistance protein 1 (MRP1), an ATP-binding cassette protein, causes multidrug resistance. We developed a functional cysteine-less version of MRP1 that provides a framework for detailed biochemical and biophysical studies. The 18 Cys residues of a truncated MRP1 (tMRP1; lacking the first multispanning transmembrane domain) were replaced with Ala to generate Cys-less tMRP1 (CL tMRP1). CL tMRP1 expressed in Saccharomyces cerevisiae membranes displayed high-affinity ATP-dependent transport of the MRP1 substrate leukotriene C4. Compared with full-length MRP1, the K m for leukotriene C4 transport by CL tMRP1 was increased approximately 3-fold, while V max was not affected. Thus a functional CL tMRP1 can be expressed using a low-cost and rapid-generation yeast expression system. This Cys-less protein can be used for biochemical, spectroscopic and structural studies to elucidate the mechanism of drug transport by MRP1.

Citing Articles

Directed evolution of P-glycoprotein cysteines reveals site-specific, non-conservative substitutions that preserve multidrug resistance.

Swartz D, Mok L, Botta S, Singh A, Altenberg G, Urbatsch I Biosci Rep. 2014; 34(3).

PMID: 24825346 PMC: 4069687. DOI: 10.1042/BSR20140062.

Effect of cysteine mutagenesis on the function and disulfide bond formation of human ABCG2.

Liu Y, Yang Y, Qi J, Peng H, Zhang J J Pharmacol Exp Ther. 2008; 326(1):33-40.

PMID: 18430864 PMC: 2632310. DOI: 10.1124/jpet.108.138115.

The remarkable transport mechanism of P-glycoprotein: a multidrug transporter.

Al-Shawi M, Omote H J Bioenerg Biomembr. 2006; 37(6):489-96.

PMID: 16691488 PMC: 1459968. DOI: 10.1007/s10863-005-9497-5.

References

Leslie E, Deeley R, Cole S . Toxicological relevance of the multidrug resistance protein 1, MRP1 (ABCC1) and related transporters. Toxicology. 2001; 167(1):3-23. DOI: 10.1016/s0300-483x(01)00454-1. View

Locher K, Lee A, Rees D . The E. coli BtuCD structure: a framework for ABC transporter architecture and mechanism. Science. 2002; 296(5570):1091-8. DOI: 10.1126/science.1071142. View

Morello J, Bichet D, Bouvier M . Pharmacological chaperones: a new twist on receptor folding. Trends Pharmacol Sci. 2000; 21(12):466-9. DOI: 10.1016/s0165-6147(00)01575-3. View

Loe D, Almquist K, Deeley R, Cole S . Multidrug resistance protein (MRP)-mediated transport of leukotriene C4 and chemotherapeutic agents in membrane vesicles. Demonstration of glutathione-dependent vincristine transport. J Biol Chem. 1996; 271(16):9675-82. DOI: 10.1074/jbc.271.16.9675. View

Ambudkar S, Dey S, Hrycyna C, Ramachandra M, Pastan I, Gottesman M . Biochemical, cellular, and pharmacological aspects of the multidrug transporter. Annu Rev Pharmacol Toxicol. 1999; 39:361-98. DOI: 10.1146/annurev.pharmtox.39.1.361. View

Renes J, de Vries E, Jansen P, Muller M . The (patho)physiological functions of the MRP family. Drug Resist Updat. 2001; 3(5):289-302. DOI: 10.1054/drup.2000.0156. View

Leier I, Jedlitschky G, Buchholz U, Cole S, Deeley R, Keppler D . The MRP gene encodes an ATP-dependent export pump for leukotriene C4 and structurally related conjugates. J Biol Chem. 1994; 269(45):27807-10. View

Bakos E, Evers R, Szakacs G, Tusnady G, Welker E, Szabo K . Functional multidrug resistance protein (MRP1) lacking the N-terminal transmembrane domain. J Biol Chem. 1998; 273(48):32167-75. DOI: 10.1074/jbc.273.48.32167. View

Higgins C, Linton K . Structural biology. The xyz of ABC transporters. Science. 2001; 293(5536):1782-4. DOI: 10.1126/science.1065588. View

10.

Yang Y, Chen Q, Zhang J . Structural and functional consequences of mutating cysteine residues in the amino terminus of human multidrug resistance-associated protein 1. J Biol Chem. 2002; 277(46):44268-77. DOI: 10.1074/jbc.M207003200. View

11.

Loo T, Clarke D . Determining the structure and mechanism of the human multidrug resistance P-glycoprotein using cysteine-scanning mutagenesis and thiol-modification techniques. Biochim Biophys Acta. 1999; 1461(2):315-25. DOI: 10.1016/s0005-2736(99)00165-0. View

12.

Zaman G, Flens M, van Leusden M, de Haas M, Mulder H, Lankelma J . The human multidrug resistance-associated protein MRP is a plasma membrane drug-efflux pump. Proc Natl Acad Sci U S A. 1994; 91(19):8822-6. PMC: 44698. DOI: 10.1073/pnas.91.19.8822. View

13.

Figler R, Omote H, Nakamoto R . Use of chemical chaperones in the yeast Saccharomyces cerevisiae to enhance heterologous membrane protein expression: high-yield expression and purification of human P-glycoprotein. Arch Biochem Biophys. 2000; 376(1):34-46. DOI: 10.1006/abbi.2000.1712. View

14.

Han E, Vanoye C, Altenberg G, Reuss L . P-glycoprotein-associated chloride currents revealed by specific block by an anti-P-glycoprotein antibody. Am J Physiol. 1996; 270(5 Pt 1):C1370-8. DOI: 10.1152/ajpcell.1996.270.5.C1370. View

15.

Borst P, Evers R, Kool M, Wijnholds J . The multidrug resistance protein family. Biochim Biophys Acta. 1999; 1461(2):347-57. DOI: 10.1016/s0005-2736(99)00167-4. View

16.

Chang G, Roth C . Structure of MsbA from E. coli: a homolog of the multidrug resistance ATP binding cassette (ABC) transporters. Science. 2001; 293(5536):1793-800. DOI: 10.1126/science.293.5536.1793. View

17.

Cai J, Daoud R, Georges E, Gros P . Functional expression of multidrug resistance protein 1 in Pichia pastoris. Biochemistry. 2001; 40(28):8307-16. DOI: 10.1021/bi010093c. View

18.

Hipfner D, Deeley R, Cole S . Structural, mechanistic and clinical aspects of MRP1. Biochim Biophys Acta. 1999; 1461(2):359-76. DOI: 10.1016/s0005-2736(99)00168-6. View

19.

Davidson A . Structural biology. Not just another ABC transporter. Science. 2002; 296(5570):1038-40. DOI: 10.1126/science.1072484. View