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

被引:12
作者
Lee, SH
Altenberg, GA [1 ]
机构
[1] Univ Texas, Med Branch, Membrane Prot Lab, Sealy Ctr Struct Biol, Galveston, TX 77555 USA
[2] Univ Texas, Med Branch, Dept Physiol & Biophys, Galveston, TX 77555 USA
关键词
ATP-binding cassette (ABC) protein; Cys-less; cystic fibrosis transmembrane conductance regulator (CFTR); MRP; multidrug resistance; nucleotide-binding domain;
D O I
10.1042/BJ20021452
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Overexpression of multidrug resistance protein I (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 C-4. Compared with full-length MRP1, the K-m for leukotriene C-4 transport by CL tMRP1 was increased similar to3-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.
引用
收藏
页码:357 / 360
页数:4
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