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. 2003 Feb 15;370(Pt 1):357–360. doi: 10.1042/BJ20021452

Transport of leukotriene C4 by a cysteine-less multidrug resistance protein 1 (MRP1).

Sung H Lee 1, Guillermo A Altenberg 1
PMCID: PMC1223133  PMID: 12398766

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.

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Selected References

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Supplementary Materials

Details on plasmid engineering
bj3700357add.pdf (174.3KB, pdf)

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