scholarly journals Mutational Disruption of Plasma Membrane Trafficking of Saccharomyces cerevisiae Yor1p, a Homologue of Mammalian Multidrug Resistance Protein

1999 ◽  
Vol 19 (4) ◽  
pp. 2998-3009 ◽  
Author(s):  
David J. Katzmann ◽  
Eric A. Epping ◽  
W. Scott Moye-Rowley
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Plasma Membrane ◽  
Multidrug Resistance ◽  
Abc Transporter ◽  
Membrane Trafficking ◽  
Predicted Amino Acid Sequence ◽  
Multidrug Resistance Protein ◽  
Wild Type ◽  
Amino Terminal ◽  
Resistance Protein

ABSTRACT The ATP binding cassette (ABC) transporter protein Yor1p was identified on the basis of its ability to elevate oligomycin resistance when it was overproduced from a high-copy-number plasmid. Analysis of the predicted amino acid sequence of Yor1p indicated that this protein was a new member of a subfamily of ABC transporter proteins defined by the multidrug resistance protein (MRP). In this work, Yor1p is demonstrated to localize to the Saccharomyces cerevisiaeplasma membrane by both indirect immunofluorescence and biochemical fractionation studies. Several mutations were generated in the amino-terminal nucleotide binding domain (NBD1) of Yor1p to test if the high degree of sequence conservation in this region of the protein was important for function. Deletion of a phenylalanine residue at Yor1p position 670 led to a mutant protein that appeared to be retained in the endoplasmic reticulum (ER) and that was unstable. As shown by others, deletion of the analogous residue from a second mammalian MRP family member, the cystic fibrosis transmembrane conductance regulator (CFTR), also led to retention of this normally plasma membrane-localized protein in the ER. Changes in the spacing between or the sequences flanking functional motifs of Yor1p NBD1 led to defective trafficking or decreased activity of the mutant proteins. Analyses of the degradation of wild-type and ΔF670 Yor1p indicated that the half-life of ΔF670 Yor1p was dramatically shortened. While the vacuole was the primary site for turnover of wild-type Yor1p, degradation of ΔF670 Yor1p was found to be more complex with both proteasomal and vacuolar contributions.

The human multidrug resistance protein MRP1 translocates sphingolipid analogs across the plasma membrane

1999 ◽  
Vol 112 (3) ◽  
pp. 415-422 ◽  
Author(s):  
R.J. Raggers ◽  
A. van Helvoort ◽  
R. Evers ◽  
G. van Meer
Keyword(s):  
Plasma Membrane ◽  
Multidrug Resistance ◽  
Abc Transporter ◽  
Basolateral Membrane ◽  
Brefeldin A ◽  
Apical Plasma Membrane ◽  
Multidrug Resistance Protein ◽  
Vesicular Traffic ◽  
P Glycoprotein ◽  
Resistance Protein

Recently, we have provided evidence that the ABC-transporter MDR1 P-glycoprotein translocates analogs of various lipid classes across the apical plasma membrane of polarized LLC-PK1 cells transfected with MDR1 cDNA. Here, we show that expression of the basolateral ABC-transporter MRP1 (the multidrug resistance protein) induced lipid transport to the exoplasmic leaflet of the basolateral plasma membrane of LLC-PK1 cells at 15 degreesC. C6-NBD-glucosylceramide synthesized on the cytosolic side of the Golgi complex, but not C6-NBD-sphingomyelin synthesized in the Golgi lumen, became accessible to depletion by BSA in the basal culture medium. This suggests the absence of vesicular traffic and direct translocation of C6-NBD-glucosylceramide by MRP1 across the basolateral membrane. In line with this, transport of the lipid to the exoplasmic leaflet depended on the intracellular glutathione concentration and was inhibited by the MRP1-inhibitors sulfinpyrazone and indomethacin, but not by the MDR1 P-glycoprotein inhibitor PSC 833. In contrast to the broad substrate specificity of the MDR1 P-glycoprotein, MRP1 selectively transported C6-NBD-glucosylceramide and C6-NBD-sphingomyelin, the latter only when it was released from the Golgi lumen by brefeldin A. This shows the specific nature of the lipid translocation. We conclude that the transport activity of MDR1 P-glycoprotein and MRP1 must be taken into account in studies on the transport of lipids to the cell surface.

scholarly journals Genetic variation of the ABC transporter gene ABCC1 (Multidrug resistance protein 1 – MRP1) in the Polish population

BMC Genetics ◽  
2015 ◽  
Vol 16 (1) ◽  
Author(s):  
Marcin Słomka ◽  
Marta Sobalska-Kwapis ◽  
Małgorzata Korycka-Machała ◽  
Grzegorz Bartosz ◽  
Jarosław Dziadek ◽  
...  
Keyword(s):  
Genetic Variation ◽  
Multidrug Resistance ◽  
Abc Transporter ◽  
Multidrug Resistance Protein ◽  
Polish Population ◽  
Transporter Gene ◽  
Multidrug Resistance Protein 1 ◽  
Resistance Protein

Breast Cancer Resistance Protein and Multidrug Resistance Protein 2 Mediate the Disposition of Leonurine-10-O-β-glucuronide

2020 ◽  
Vol 21 (13) ◽  
pp. 1060-1067
Author(s):  
Xiaocui Li ◽  
Yushan Xie ◽  
Wei Qu ◽  
Xiaojun Ou ◽  
Xiaowen Ou ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Multidrug Resistance ◽  
Breast Cancer Resistance Protein ◽  
Gene Knockout ◽  
Multidrug Resistance Protein ◽  
Cancer Resistance ◽  
Wild Type ◽  
Multidrug Resistance Protein 2 ◽  
Intestinal Microsomes ◽  
Resistance Protein

Background: Leonurine (Leo), a promising antilipemic agent that has been approved for clinical trials, is extensively metabolized into bioactive Leonurine-10-O-β-glucuronide (L-10-G) vivo. Objective: To explore the effects of breast cancer resistance protein (Bcrp) and multidrug resistance protein 2 (Mrp2) on the disposition of L-10-G. Methods: The pharmacokinetics, tissue distribution and intestinal perfusion of Leo were studied by using efflux transporter gene knockout mouse models. The enzyme kinetics via liver and intestinal microsomes were also examined. Results: After intravenous injection with Leo, the AUC0-∞ values of L-10-G in Bcrp1-/- and Mrp2-/- mice were 1.55-fold and 16.80-fold higher, respectively, than those in wild-type FVB mice (P < 0.05). After oral administration, the AUC0-∞ value of L-10-G showed a 2.82-fold increase in Mrp2-/- mice compared with wild-type FVB mice (P < 0.05). After gavage with Leo for 10 and 25 min, the bile accumulation of L-10-G in Mrp2-/- mice was 3-fold and 22-fold lower, respectively, than that in wild-type FVB mice (P < 0.05). Besides, the intestinal excreted amount of L-10-G showed 2.22-fold and 2.68-fold decrease in Bcrp1-/- and Mrp2-/- mice, respectively, compared with that in wild-type FVB mice (P < 0.05). The clearance of L-10-G decreased in liver microsomes and increased in intestinal microsomes of Bcrp1-/- and Mrp2-/- mice compared to the wild-type FVB mice (P < 0.05). Conclusion: Both Bcrp and Mrp2 are involved in the disposition of L-10-G, and Mrp2 exhibits a superior influence.

Characterization of the amino-terminal regions in the human multidrug resistance protein (MRP1)

2000 ◽  
Vol 113 (24) ◽  
pp. 4451-4461
Author(s):  
E. Bakos ◽  
R. Evers ◽  
G. Calenda ◽  
G.E. Tusnady ◽  
G. Szakacs ◽  
...  
Keyword(s):  
Multidrug Resistance ◽  
Drug Transport ◽  
Cellular Localization ◽  
Membrane Vesicles ◽  
Sf9 Cells ◽  
Multidrug Resistance Protein ◽  
Hydrophobic Membrane ◽  
Amino Terminal ◽  
Mdckii Cells ◽  
Resistance Protein

The human multidrug resistance protein (MRP1) contributes to drug resistance in cancer cells. In addition to an MDR1-like core, MRP1 contains an N-terminal membrane-bound (TMD(0)) region and a cytoplasmic linker (L(0)), both characteristic of several members of the MRP family. In order to study the role of the TMD(0) and L(0) regions, we constructed various truncated and mutated MRP1, and chimeric MRP1-MDR1 molecules, which were expressed in insect (Sf9) and polarized mammalian (MDCKII) cells. The function of the various proteins was examined in isolated membrane vesicles by measuring the transport of leukotriene C(4) and other glutathione conjugates, and by vanadate-dependent nucleotide occlusion. Cellular localization, and glutathione-conjugate and drug transport, were also studied in MDCKII cells. We found that chimeric proteins consisting of N-terminal fragments of MRP1 fused to the N terminus of MDR1 preserved the transport, nucleotide occlusion and apical membrane routing of wild-type MDR1. As shown before, MRP1 without TMD(0)L(0) (Delta MRP1), was non-functional and localized intracellularly, so we investigated the coexpression of Delta MRP1 with the isolated L(0) region. Coexpression yielded a functional MRP1 molecule in Sf9 cells and routing to the lateral membrane in MDCKII cells. Interestingly, the L(0) peptide was found to be associated with membranes in Sf9 cells and could only be solubilized by urea or detergent. A 10-amino-acid deletion in a predicted amphipathic region of L(0) abolished its attachment to the membrane and eliminated MRP1 transport function, but did not affect membrane routing. Taken together, these experiments suggest that the L(0) region forms a distinct domain within MRP1, which interacts with hydrophobic membrane regions and with the core region of MRP1.

Substitution of Trp1242 of TM17 alters substrate specificity of human multidrug resistance protein 3

2003 ◽  
Vol 284 (2) ◽  
pp. G280-G289 ◽  
Author(s):  
Curtis J. Oleschuk ◽  
Roger G. Deeley ◽  
Susan P. C. Cole
Keyword(s):  
Multidrug Resistance ◽  
Substrate Specificity ◽  
Multidrug Resistance Protein ◽  
Wild Type ◽  
Multidrug Resistance Protein 3 ◽  
17Β Estradiol ◽  
Resistance Protein ◽  
Estradiol 17Β ◽  
Taurocholate Transport

Multidrug resistance protein 3 (MRP3) is an ATP-dependent transporter of 17β-estradiol 17β(d-glucuronide) (E217βG), leukotriene C4 (LTC4), methotrexate, and the bile salts taurocholate and glycocholate. In the present study, the role of a highly conserved Trp residue at position 1242 on MRP3 transport function was examined by expressing wild-type MRP3 and Ala-, Cys-, Phe-, Tyr-, and Pro-substituted mutants in human embryonic kidney 293T cells. Four MRP3-Trp1242 mutants showed significantly increased E217βG uptake, whereas transport by the Pro mutant was undetectable. Similarly, the Pro mutant did not transport LTC4. By comparison, LTC4transport by the Ala, Cys, Phe, and Tyr mutants was reduced by ∼35%. The Ala, Cys, Phe, and Tyr mutants all showed greatly reduced methotrexate and leucovorin transport, except the Tyr mutant, which transported leucovorin at levels comparable with wild-type MRP3. In contrast, the MRP3-Trp1242 substitutions did not significantly affect taurocholate transport or taurocholate and glycocholate inhibition of E217βG uptake. Thus Trp1242 substitutions markedly alter the substrate specificity of MRP3 but leave bile salt binding and transport intact.

scholarly journals Complex Polymorphisms in the Plasmodium falciparum Multidrug Resistance Protein 2 Gene and Its Contribution to Antimalarial Response

2014 ◽  
Vol 58 (12) ◽  
pp. 7390-7397 ◽  
Author(s):  
Maria Isabel Veiga ◽  
Nuno S. Osório ◽  
Pedro Eduardo Ferreira ◽  
Oscar Franzén ◽  
Sabina Dahlstrom ◽  
...  
Keyword(s):  
Plasmodium Falciparum ◽  
Multidrug Resistance ◽  
Abc Transporter ◽  
Parasite Clearance ◽  
Antimalarial Drugs ◽  
Multidrug Resistance Protein ◽  
Content Type ◽  
Resistance Protein

ABSTRACTPlasmodium falciparumhas the capacity to escape the actions of essentially all antimalarial drugs. ATP-binding cassette (ABC) transporter proteins are known to cause multidrug resistance in a large range of organisms, including theApicomplexaparasites.P. falciparumgenome analysis has revealed two genes coding for the multidrug resistance protein (MRP) type of ABC transporters:Pfmrp1, previously associated with decreased parasite drug susceptibility, and the poorly studiedPfmrp2. The role ofPfmrp2polymorphisms in modulating sensitivity to antimalarial drugs has not been established. We herein report a comprehensive account of thePfmrp2genetic variability in 46 isolates from Thailand. A notably high frequency of 2.8 single nucleotide polymorphisms (SNPs)/kb was identified for this gene, including some novel SNPs. Additionally, we found thatPfmrp2harbors a significant number of microindels, some previously not reported. We also investigated the potential association of the identifiedPfmrp2polymorphisms with alteredin vitrosusceptibility to several antimalarials used in artemisinin-based combination therapy and with parasite clearance time. Association analysis suggestedPfmrp2polymorphisms modulate the parasite'sin vitroresponse to quinoline antimalarials, including chloroquine, piperaquine, and mefloquine, and association within vivoparasite clearance. In conclusion, our study reveals that thePfmrp2gene is the most diverse ABC transporter known inP. falciparumwith a potential role in antimalarial drug resistance.

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