Expression and Function of Human MRP1 (ABCC1) Is Dependent on Amino Acids in Cytoplasmic Loop 5 and Its Interface with Nucleotide Binding Domain 2

Iram, Surtaj H.; Cole, Susan P.C.

doi:10.1074/jbc.m110.166959

Cited by 30 publications

(47 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…MRP1 is a glycoprotein known as ABCC1 and belongs to the ATP-binding protein superfamily [8] . This molecule has three membrane-spanning domains and two nucleotide-binding domains, which function as the ATP-dependent drug pump that effluxes drugs and organic anions across the plasma membrane [4,9] .…”

Section: Discussionmentioning

confidence: 99%

Multidrug resistance-associated protein 1 decreases the concentrations of antiepileptic drugs in cortical extracellular fluid in amygdale kindling rats

et al. 2013

View full text Add to dashboard Cite

Aim: To investigate whether multidrug resistance-associated protein 1 (MRP1) was responsible for drug resistence in refractory epilepsy in amygdale kindling rats. Methods: Rat amygdale kindling was used as a model of refractory epilepsy. The expression of MRP1 mRNA and protein in the brains was examined using RT-PCR and Western blot. MRP1-positive cells in the cortex and hippocampus were studied with immunohistochemical staining. The rats were intraperitoneally injected with phenytoin (50 mg/kg) or carbamazepine (20 mg/kg), and their concentrations in the cortical extracellular fluid were measured using microdialysis and HPLC. Probenecid, a MRP1 inhibitor (40 mmol/L, 50 μL) was administered through an inflow tube into the cortex 30 min before injection of the antiepileptic drugs. Results: The expression of MRP1 mRNA and protein was significantly up-regulated in the cortex and hippocampus in amygdale kindling rats compared with the control group. Furthermore, the number of MRP1-positive cells in the cortex and hippocampus was also significantly increased in amygdale kindling rats. Microdialysis studies showed that the concentrations of phenytoin and carbamazepine in the cortical extracellular fluid were significantly decreased in amygdale kindling rats. Pre-administration of probenecid could restore the concentrations back to their control levels. Conclusion: Up-regulation of MRP1 is responsible for the resistance of brain cells to antiepileptic drugs in the amygdale kindling rats.

show abstract

Section: Discussionmentioning

confidence: 99%

Multidrug resistance-associated protein 1 decreases the concentrations of antiepileptic drugs in cortical extracellular fluid in amygdale kindling rats

et al. 2013

View full text Add to dashboard Cite

show abstract

“…The generation of wild-type, K513A, K516A, E521A, and E535A mutant MRP1 pcDNA3.1 expression vectors has been described previously (19,21). The expression vectors (20 g of DNA) were transfected into 90 -95% confluent SV40-transformed human embryonic kidney cells (HEK293T) using Lipofectamine 2000 (Invitrogen).…”

Section: Transfections Preparation Of Membrane Vesicles and Measurementioning

confidence: 99%

“…Inspection of homology models of MRP1 showed that three of the four latter charged residues are predicted to lie at the interface of CL5 with NBD2 (see Fig. 1B) (11,19), and selected mutations in NBD2 allowed us to conclude that bonding interactions between CL5 and NBD2 play a pivotal role in the proper folding and assembly and, hence, expression of MRP1 at the plasma membrane. Our findings also demonstrated that interactions of the fourth amino acid, Glu 535 , with certain amino acids in CL5 and CL6 were important for efficient expression of MRP1 in mammalian cells.…”

mentioning

confidence: 99%

“…1, A and B) (19). Our earlier studies showed that CL5, which is more highly conserved than CL7, plays a greater role in supporting proper membrane expression and function of MRP1.…”

mentioning

confidence: 99%

“…Our earlier studies showed that CL5, which is more highly conserved than CL7, plays a greater role in supporting proper membrane expression and function of MRP1. Thus, mutational analysis of eight conserved charged amino acids in CL5 revealed that four of them (Arg 501 , Glu 507 , Arg 532 , Gly 511 ) are important for MRP1 transport activity, whereas the other four (Lys 513 , Lys 516 , Glu 521 , Glu 535 ) are essential for plasma membrane expression of the fully glycosylated MRP1 protein in human embryonic kidney (HEK) cells (19). Inspection of homology models of MRP1 showed that three of the four latter charged residues are predicted to lie at the interface of CL5 with NBD2 (see Fig.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Mutation of Glu521 or Glu535 in Cytoplasmic Loop 5 Causes Differential Misfolding in Multiple Domains of Multidrug and Organic Anion Transporter MRP1 (ABCC1)

Iram

Cole

2012

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Background:The five-domain MRP1 mediates ATP-dependent efflux of drugs and organic anions. Results: Chemically rescued cytoplasmic loop 5 (CL5) processing mutants of MRP1 exhibit distinct phenotypes characterized by differences in transport activity and protein conformation. Conclusion: Glu 521 and Glu 535 are important for folding of their resident domain as well as more COOH-proximal domains. Significance: MRP1 folding processes differ from those of its homologs including CFTR.

show abstract

Study on functional sites in human multidrug resistance protein 1 (hMRP1)

Han

Farooq

et al. 2021

Proteins

View full text Add to dashboard Cite

Human multidrug resistance protein 1 (hMRP1) is an important member of the ATPbinding cassette (ABC) transporter superfamily. It can extrude a variety of anticancer drugs and physiological organic anions across the plasma membrane, which is activated by substrate binding, and is accompanied by large-scale cooperative movements between different domains. Currently, it remains unclear completely about how the specific interactions between hMRP1 and its substrate are and which critical residues are responsible for allosteric signal transduction. To the end, we first construct an inward-facing state of hMRP1 using homology modeling method, and then dock substrate proinflammatory agent leukotriene C4 (LTC4) to hMRP1 pocket. The result manifests LTC4 interacts with two parts of hMRP1 pocket, namely the positively charged pocket (P pocket) and hydrophobic pocket (H pocket), similar to its binding mode with bMRP1 (bovine MRP1). Additionally, we use the Gaussian network model (GNM)-based thermodynamic method proposed by us to identify the key residues whose perturbations markedly alter their binding free energy. Here the conventional GNM is improved with covalent/non-covalent interactions and secondary structure information considered (denoted as sscGNM). In the result, sscGNM improves the flexibility prediction, especially for the nucleotide binding domains with rich kinds of secondary structures. The 46 key residue clusters located in different subdomains are identified which are highly consistent with experimental observations. Furtherly, we explore the long-range cooperation within the transporter. This study is helpful for strengthening the understanding of the work mechanism in ABC exporters and can provide important information to scientists in drug design studies.binding mode, Gaussian network model, hMRP1, key residues, thermodynamic cycle | INTRODUCTIONMultidrug resistance (MDR) to chemotherapy is a major obstacle in the treatment of cancer patients. Human multidrug resistance protein 1 (hMRP1), a member of the ATP-binding cassette (ABC) transporter superfamily, can export a wide variety of anticancer drugs such as vincristine, etoposide, anthracyclines, methotrexate, and physiological organic anions antioxidants GSH and proinflammatory agent leukotriene C4 (LTC4) across the cell membrane utilizing the energy from ATP binding and hydrolysis. 1 Studies have shown that the process of substrate transport is accompanied by large-scale cooperative motions between spatially separated subdomains. 2 Thus, there must exist a network of key residues mediating long-range signal transmission and allosteric coupling. 2,3 Identification of key functional residues is very important not only for an "in-depth" understanding of the mechanism of signal transmission but also for the structure-based drug design.

show abstract

Expression and Function of Human MRP1 (ABCC1) Is Dependent on Amino Acids in Cytoplasmic Loop 5 and Its Interface with Nucleotide Binding Domain 2

Cited by 30 publications

References 48 publications

Multidrug resistance-associated protein 1 decreases the concentrations of antiepileptic drugs in cortical extracellular fluid in amygdale kindling rats

Multidrug resistance-associated protein 1 decreases the concentrations of antiepileptic drugs in cortical extracellular fluid in amygdale kindling rats

Mutation of Glu521 or Glu535 in Cytoplasmic Loop 5 Causes Differential Misfolding in Multiple Domains of Multidrug and Organic Anion Transporter MRP1 (ABCC1)

Study on functional sites in human multidrug resistance protein 1 (hMRP1)

Contact Info

Product

Resources

About