ABCC1 p.Lys1141Arg
| Predicted by SNAP2: | A: D (80%), C: D (85%), D: D (95%), E: D (91%), F: D (85%), G: D (91%), H: D (85%), I: D (85%), L: D (85%), M: D (71%), N: D (91%), P: D (91%), Q: D (80%), R: D (59%), S: D (85%), T: D (85%), V: D (85%), W: D (91%), Y: D (85%), |
| Predicted by PROVEAN: | A: D, C: D, D: D, E: D, F: D, G: D, H: D, I: D, L: D, M: D, N: D, P: D, Q: D, R: N, S: D, T: D, V: D, W: D, Y: D, |
[switch to compact view]
Comments [show]
None has been submitted yet.
[hide] Functional importance of three basic residues clus... J Biol Chem. 2006 Jan 6;281(1):43-50. Epub 2005 Oct 17. Conseil G, Deeley RG, Cole SP
Functional importance of three basic residues clustered at the cytosolic interface of transmembrane helix 15 in the multidrug and organic anion transporter MRP1 (ABCC1).
J Biol Chem. 2006 Jan 6;281(1):43-50. Epub 2005 Oct 17., 2006-01-06 [PMID:16230346]
Abstract [show]
The multidrug resistance protein 1 (MRP1) mediates drug and organic anion efflux across the plasma membrane. The 17 transmembrane (TM) helices of MRP1 are linked by extracellular and cytoplasmic (CL) loops of various lengths and two cytoplasmic nucleotide binding domains. In this study, three basic residues clustered at the predicted TM15/CL7 interface were investigated for their role in MRP1 expression and activity. Thus, Arg1138, Lys1141, and Arg1142 were replaced with residues of the same or opposite charge, expressed in human embryonic kidney cells, and the properties of the mutant proteins were assessed. Neither Glu nor Lys substitutions of Arg1138 and Arg1142 affected MRP1 expression; however, all four mutants showed a decrease in organic anion transport with a relatively greater decrease in leukotriene C4 and glutathione transport. These mutations also modulated MRP1 ATPase activity as reflected by a decreased vanadate-induced trapping of 8-azido-[32P]ADP. Mutation of Lys1141 to either Glu or Arg reduced MRP1 expression, and routing to the plasma membrane was impaired. However, only the Glu-substituted Lys1141 mutant showed a decrease in organic anion transport, and this was associated with decreased substrate binding and vanadate-induced trapping of 8-azido-ADP. These studies identified a cluster of basic amino acids likely at the TM15/CL7 interface as a region important for both MRP1 expression and activity and demonstrated that each of the three residues plays a distinct role in the substrate specificity and catalytic activity of the transporter.
Comments [show]
None has been submitted yet.
No. Sentence Comment
6 Mutation of Lys1141 to either Glu or Arg reduced MRP1 expression, and routing to the plasma membrane was impaired.
X
ABCC1 p.Lys1141Arg 16230346:6:12
status: NEW47 Substitutions of charged residues were generated in the pGEM-3Z- XmaI/MRP1 plasmid according to the manufacturer`s instructions with the following mutagenic primers (Integrated DNA Technologies, Inc., Coralville, IA), which also introduced an additional silent restriction site (substituted nucleotides are underlined): R1138E, 5Ј-CGT GGC TTC CTC CGA GCA ACT GAA GCG CCT CGA G-3Ј; R1138K, 5Ј-CGT GGC TTC CTC CAA GCA GCT TAA GCG CCT CGA G-3Ј; K1141E, 5Ј-CGG CAG CTG GAG CGC CTG GAG TCG GTC AGC-3Ј; K1141R, 5Ј-CCC GGC AGC TGC GGC GCC TCG AGT CGG-3Ј; R1142E, 5Ј-CCC GGC AGC TGA AGG AGC TCG AGT CGG TCA GCC G-3Ј;R1142K, 5Ј-CCC GGC AGC TGA AGA AGC TTG AGT CGG TCA GCC G-3Ј.
X
ABCC1 p.Lys1141Arg 16230346:47:533
status: NEW122 B, HEK293T cells were transfected with expression vectors encoding WT-MRP1 and MRP1 mutants R1138E, R1138K, K1141E, K1141R, R1142E, and R1142K, and 48 h later, cells were probed with monoclonal antibody QCRL-3 and processed for confocal fluorescence microscopy using Alexa Fluor 488 conjugated secondary antibody.
X
ABCC1 p.Lys1141Arg 16230346:122:116
status: NEW141 Kinetic parameters (Vmax and Km) were obtained after plotting the data according to the Michaelis-Menten equation for wild-type MRP1 and the K1141E and K1141R mutants (Table 1).
X
ABCC1 p.Lys1141Arg 16230346:141:152
status: NEW143 However, although the Vmax for the K1141E mutant was unchanged, its Km for E217betaG transport was significantly increased by 4.4-fold (8.8 Ϯ 0.7 M) when compared with the K1141R mutant (2.0 Ϯ 0.3 M) and wild-type MRP1 (1.9 Ϯ 0.4 M).
X
ABCC1 p.Lys1141Arg 16230346:143:186
status: NEW160 TABLE 1 Kinetic parameters of vesicular uptake of E2 17betaG by wild-type and Lys1141 mutant MRP1 Km Vmax a Vmax/Km ؋ 103 M pmol mg-1 min-1 mg/l/min WT-MRP1 1.9 Ϯ 0.4 604 Ϯ 36 0.32 K1141E 8.8 Ϯ 0.7 577 Ϯ 20 0.07 K1141R 2.0 Ϯ 0.3 964 Ϯ 42 0.48 a Vmax values have been corrected for differences in protein expression of the Lys1141 mutants relative to WT-MRP1 (Fig. 2A).
X
ABCC1 p.Lys1141Arg 16230346:160:250
status: NEW165 Consistent with this interpretation, the orthovanadate-induced azido-ADP trapping by the same charge K1141R mutant, which had the least affected transport activity of all the mutants, was comparable with wild-type MRP1 (ϳ90%).
X
ABCC1 p.Lys1141Arg 16230346:165:101
status: NEW204 Nevertheless, the vesicular transport activity of the K1141R mutant was similar to wild-type MRP1, as was its ability to be photolabeled with LTC4 and its ATPase activity.
X
ABCC1 p.Lys1141Arg 16230346:204:54
status: NEW[hide] Structural and functional properties of human mult... Curr Med Chem. 2011;18(3):439-81. He SM, Li R, Kanwar JR, Zhou SF
Structural and functional properties of human multidrug resistance protein 1 (MRP1/ABCC1).
Curr Med Chem. 2011;18(3):439-81., [PMID:21143116]
Abstract [show]
Multidrug ABC transporters such as P-glycoprotein (P-gp/MDR1/ABCB1) and multidrug resistance protein 1 (MRP1/ABCC1) play an important role in the extrusion of drugs from the cell and their overexpression can be a cause of failure of anticancer and antimicrobial chemotherapy. Recently, the mouse P-gp/Abcb1a structure has been determined and this has significantly enhanced our understanding of the structure-activity relationship (SAR) of mammalian ABC transporters. This paper highlights our current knowledge on the structural and functional properties and the SAR of human MRP1/ABCC1. Although the crystal structure of MRP1/ABCC1 has yet to be resolved, the current topological model of MRP1/ABCC1 contains two transmembrane domains (TMD1 and TMD2) each followed by a nucleotide binding domain (NBD) plus a third NH2-terminal TMD0. MRP1/ABCC1 is expressed in the liver, kidney, intestine, brain and other tissues. MRP1/ABCC1 transports a structurally diverse array of important endogenous substances (e.g. leukotrienes and estrogen conjugates) and xenobiotics and their metabolites, including various conjugates, anticancer drugs, heavy metals, organic anions and lipids. Cells that highly express MRP1/ABCC1 confer resistance to a variety of natural product anticancer drugs such as vinca alkaloids (e.g. vincristine), anthracyclines (e.g. etoposide) and epipodophyllotoxins (e.g. doxorubicin and mitoxantrone). MRP1/ABCC1 is associated with tumor resistance which is often caused by an increased efflux and decreased intracellular accumulation of natural product anticancer drugs and other anticancer agents. However, most compounds that efficiently reverse P-gp/ABCB1-mediated multidrug resistance have only low affinity for MRP1/ABCC1 and there are only a few effective and relatively specific MRP1/ABCC1 inhibitors available. A number of site-directed mutagenesis studies, biophysical and photolabeling studies, SAR and QSAR, molecular docking and homology modeling studies have documented the role of multiple residues in determining the substrate specificity and inhibitor selectivity of MRP1/ABCC1. Most of these residues are located in the TMs of TMD1 and TMD2, in particular TMs 4, 6, 7, 8, 10, 11, 14, 16, and 17, or in close proximity to the membrane/cytosol interface of MRP1/ABCC1. The exact transporting mechanism of MRP1/ABCC1 is unclear. MRP1/ABCC1 and other multidrug transporters are front-line mediators of drug resistance in cancers and represent important therapeutic targets in future chemotherapy. The crystal structure of human MRP1/ABCC1 is expected to be resolved in the near future and this will provide an insight into the SAR of MRP1/ABCC1 and allow for rational design of anticancer drugs and potent and selective MRP1/ABCC1 inhibitors.
Comments [show]
None has been submitted yet.
No. Sentence Comment
742 Notably, mutation of Lys1141 to either Glu or Arg impaired MRP1/ABCC1 expression and routing to the plasma membrane [354].
X
ABCC1 p.Lys1141Arg 21143116:742:21
status: NEW743 Substitution of Lys1141 with Glu (negatively chages) at TM15/CL7 interface decreased LTC4 transport by 70%, but an Arg substitution (positively charged) at this position did not; GSH transport by both Lys1141Glu and Lys1141Arg mutants was decreased by 40%, suggesting that the presence of a positive charge at Lys1141 was sufficient for LTC4 transport, whereas the less bulky side chain of Lys itself seemed important for GSH transport [354].
X
ABCC1 p.Lys1141Arg 21143116:743:216
status: NEW745 All mutants except Lys1141Arg showed substantially reduced trapping of 8-azido-[ 32P]ATP (40-60% that of wild-type MRP1), implicating that the ability of these mutants at TM15/CL7 interface to hydrolyze ATP is impaired and consequently disrupt the transport cycle and cause a decrease in transport activity.
X
ABCC1 p.Lys1141Arg 21143116:745:19
status: NEW