TY - JOUR
T1 - Characterization of the cellular localization, expression level, and function of SNP variants of MRP2/ABCC2
AU - Hirouchi, Masakazu
AU - Suzuki, Hiroshi
AU - Itoda, Masaya
AU - Ozawa, Shogo
AU - Sawada, Jun Ichi
AU - Ieiri, Ichiro
AU - Ohtsubo, Kenji
AU - Sugiyama, Yuichi
N1 - Funding Information:
This work was supported by Grant-in-Aid for Scientific Research on Priority Areas epithelial vectorial transport 12144201 from the Ministry of Education, Science and Culture of Japan, and by Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.
PY - 2004/5
Y1 - 2004/5
N2 - Purpose. The presence of single nucleotide polymorphisms (SNPs) has been reported for multidrug resistance-associated protein 2 (MRP2/ABCC2). The purpose of the current study was to characterize the localization, expression level, and function of MRP2 variants. Methods. The expression and cellular localization of the wild-type and three kinds of reported SNP variants of MRP2 molecules were analyzed in LLC-PK1 cells after infection with the recombinant Tetoff adenoviruses. Their function was determined by using the isolated membrane vesicles from the infected LLC-PK1 cells. Results. The transport activity for E217βG, LTC4, and DNP-SG, normalized by the expression level of MRP2, was similar between the wild-type, V417I, and A1450T MRP2s. The transport activity of S789F MRP2 was slightly higher than that of wild-type MRP2. However, the expression level of S789F and A1450T MRP2 proteins was significantly lower compared with the wild-type and V417I MRP2. In addition, although the wild-type and V417I MRP2 were exclusively localized in the apical membrane, S789F and A1450T MRP2 were located in the apical membrane and also in the intracellular compartment. Conclusions. These results suggest that the most frequently observed V417I substitution may not affect the in vivo function of MRP2, whereas the much less frequently observed S789F and A1450T may be associated with the reduced in vivo function.
AB - Purpose. The presence of single nucleotide polymorphisms (SNPs) has been reported for multidrug resistance-associated protein 2 (MRP2/ABCC2). The purpose of the current study was to characterize the localization, expression level, and function of MRP2 variants. Methods. The expression and cellular localization of the wild-type and three kinds of reported SNP variants of MRP2 molecules were analyzed in LLC-PK1 cells after infection with the recombinant Tetoff adenoviruses. Their function was determined by using the isolated membrane vesicles from the infected LLC-PK1 cells. Results. The transport activity for E217βG, LTC4, and DNP-SG, normalized by the expression level of MRP2, was similar between the wild-type, V417I, and A1450T MRP2s. The transport activity of S789F MRP2 was slightly higher than that of wild-type MRP2. However, the expression level of S789F and A1450T MRP2 proteins was significantly lower compared with the wild-type and V417I MRP2. In addition, although the wild-type and V417I MRP2 were exclusively localized in the apical membrane, S789F and A1450T MRP2 were located in the apical membrane and also in the intracellular compartment. Conclusions. These results suggest that the most frequently observed V417I substitution may not affect the in vivo function of MRP2, whereas the much less frequently observed S789F and A1450T may be associated with the reduced in vivo function.
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U2 - 10.1023/B:PHAM.0000026422.06207.33
DO - 10.1023/B:PHAM.0000026422.06207.33
M3 - Article
C2 - 15180328
AN - SCOPUS:3543004158
SN - 0724-8741
VL - 21
SP - 742
EP - 748
JO - Pharmaceutical Research
JF - Pharmaceutical Research
IS - 5
ER -