Identification of human multidrug resistance protein 1 (MRP1) mutations and characterization of a G671V substitution

被引:73
作者
Conrad, S
Kauffmann, HM
Ito, K
Deeley, RG
Cole, SPC
Schrenk, D
机构
[1] Univ Kaiserslautern, D-67663 Kaiserslautern, Germany
[2] Queens Univ, Canc Res Labs, Kingston, ON, Canada
关键词
human MRP1; genetic variations; expression analysis; membrane vesicles; transport assays; drug conjugate transport;
D O I
10.1007/s100380170017
中图分类号
Q3 [遗传学];
学科分类号
071007 ; 090102 ;
摘要
The multidrug resistance protein 1 (MRP1) belonging to the ATP-binding cassette (ABC) superfamily of transport proteins can confer resistance to multiple natural product drugs and methotrexate in human tumor cells. In addition, MRP1 is expressed in normal tissues acting as an efflux pump for glutathione, glucuronate, and sulfate conjugates and may thus influence the pharmacokinetic properties of many drugs. Using polymerase chain reaction-single-strand conformation polymorphism analysis, we screened 36 Caucasian volunteers for mutations in the coding exons of the MRP1 gene, including the adjacent intron sequences. Among several mutations found. two are expected to cause amino acid substitutions. One of these mutations (G671V) was of special interest because it is located near the first nucleotide binding domain. To determine whether this mutation caused a change in the MRP1 phenotype, a mutant MRP1 expression vector was constructed and transfected into SV40-transformed human embryonic kidney cells (HEKSV293T) and the transport properties of the mutant protein were examined. Transport of the MRP1 substrates leukotriene C-4, 17 beta -estradiol 17 beta-(D)-glucuronide, and estrone sulfate by membrane vesicles prepared from transiently transfected HEKSV293T cells was comparable to that of wild-type MRP1.
引用
收藏
页码:656 / 663
页数:8
相关论文
共 33 条
[1]  
ABBASZADEGAN MR, 1994, CANCER RES, V54, P4676
[2]   SINGLE-STEP METHOD OF RNA ISOLATION BY ACID GUANIDINIUM THIOCYANATE PHENOL CHLOROFORM EXTRACTION [J].
CHOMCZYNSKI, P ;
SACCHI, N .
ANALYTICAL BIOCHEMISTRY, 1987, 162 (01) :156-159
[3]  
COLE SPC, 1994, CANCER RES, V54, P5902
[4]   OVEREXPRESSION OF A TRANSPORTER GENE IN A MULTIDRUG-RESISTANT HUMAN LUNG-CANCER CELL-LINE [J].
COLE, SPC ;
BHARDWAJ, G ;
GERLACH, JH ;
MACKIE, JE ;
GRANT, CE ;
ALMQUIST, KC ;
STEWART, AJ ;
KURZ, EU ;
DUNCAN, AMV ;
DEELEY, RG .
SCIENCE, 1992, 258 (5088) :1650-1654
[5]   Sequencing and tissue distribution of the canine MRP2 gene compared with MRP1 and MDR1 [J].
Conrad, S ;
Viertelhaus, A ;
Orzechowski, A ;
Hoogstraate, J ;
Gjellan, K ;
Schrenk, D ;
Kauffmann, HM .
TOXICOLOGY, 2001, 156 (2-3) :81-91
[6]   Transport of glutathione prostaglandin A conjugates by the multidrug resistance protein 1 [J].
Evers, R ;
Cnubben, NHP ;
Wijnholds, J ;
van Deemter, L ;
van Bladeren, PJ ;
Borst, P .
FEBS LETTERS, 1997, 419 (01) :112-116
[7]   Comparison of the functional characteristics of the nucleotide binding domains of multidrug resistance protein 1 [J].
Gao, M ;
Cui, HR ;
Loe, DW ;
Grant, CE ;
Almquist, KC ;
Cole, SPC ;
Deeley, RG .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (17) :13098-13108
[8]   Analysis of the intron-exon organization of the human multidrug-resistance protein gene (MRP) and alternative splicing of its mRNA [J].
Grant, CE ;
Kurz, EU ;
Cole, SPC ;
Deeley, RG .
GENOMICS, 1997, 45 (02) :368-378
[9]   Monoclonal antibodies that inhibit the transport function of the 190-kDa multidrug resistance protein, MRP - Localization of their epitopes to the nucleotide-binding domains of the protein [J].
Hipfner, DR ;
Mao, QC ;
Qiu, W ;
Leslie, EM ;
Gao, M ;
Deeley, RG ;
Cole, SPC .
JOURNAL OF BIOLOGICAL CHEMISTRY, 1999, 274 (22) :15420-15426
[10]  
Hipfner DR, 1996, CANCER RES, V56, P3307