ABCC1 p.Arg1131Glu
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PMID: 15208328
[PubMed]
Situ D et al: "Mutational analysis of ionizable residues proximal to the cytoplasmic interface of membrane spanning domain 3 of the multidrug resistance protein, MRP1 (ABCC1): glutamate 1204 is important for both the expression and catalytic activity of the transporter."
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Sentence
Comment
83
Levels of expression of the R1046D and R1131E mutant proteins were determined by the immunoblotting of membrane vesicles prepared from the transfected HEK293T cells with mAb QCRL-1.
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ABCC1 p.Arg1131Glu 15208328:83:39
status: NEW87 To determine the functional consequences of replacing Arg1046 , Asp1084 , and Arg1131 with an oppositely charged amino acid, the ability of the R1046D, D1084R, and R1131E mutants to transport different organic anions was tested.
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ABCC1 p.Arg1131Glu 15208328:87:164
status: NEW88 Fig. 2, B and C, shows that uptake levels of E217betaG and LTC4, respectively, by the R1046D and R1131E mutants were moderately reduced (by 30-50%) compared with wild-type MRP1, whereas uptake by the D1084R mutant was substantially reduced by Ͼ90%.
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ABCC1 p.Arg1131Glu 15208328:88:97
status: NEW89 Similarly, levels of E1SO4 and MTX uptake by the R1046D and R1131E mutants were comparable with or moderately different from wild-type MRP1, whereas uptake of these organic anions by D1084R was substantially diminished (Յ25% of wild-type MRP1) (Fig. 2, D and E).
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ABCC1 p.Arg1131Glu 15208328:89:60
status: NEW93 A, MRP1 expression in membrane vesicles prepared from HEK293T cells transfected with empty vector [pcDNA3.1] and vector containing wild-type (WT-MRP1) and mutant (R1046D, D1084R, D1084E, and R1131E) cDNAs was determined by immunoblotting with mAb QCRL-1. Shown is a representative immunoblot of membrane vesicles (1 and 2 g of protein) from a single transfection.
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ABCC1 p.Arg1131Glu 15208328:93:191
status: NEW95 B-F, levels of 3 H-labeled organic anion uptake by the membrane vesicles shown in A were determined and corrected to take into account any differences in MRP1 protein expression (empty pcDNA3.1 vector control (open bars), wild-type MRP1 (black bars), and mutants R1046D, D1084R, D1084E, and R1131E (gray bars)).
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ABCC1 p.Arg1131Glu 15208328:95:291
status: NEW118 Immunoblots of membrane vesicle proteins prepared from cells expressing the Glu1204 mutants E1204L and E1204D were carried out as described in A. TABLE I Summary of organic anion transport activity of MRP1 mutants with substitutions of ionizable amino acids in and proximal to TM13 to TM17 of MSD3 Mutation % Wild-type MRP1 transport activitya E217betaG LTC4 E1SO4 MTX GSH TM13 R1046D 115 70 80 120 NDb TM14 D1084R Ͻ10 Ͻ10 15 25 Ͻ10 D1084E 80 20 65 90 20 TM15 R1131E 70 50 80 60 ND TM16 R1197E Ͻ10 Ͻ10 Ͻ15 Ͻ10 ND R1197K 20 Ͻ25 Ͻ20 Ͻ10 ND R1202G 115 115 75 70 ND R1202L 115 120 50 110 ND E1204L Ͻ10 50 10 110 Ͻ25 E1204D 100 115 100 115 Ͻ25 TM17 R1249D Ͻ10 Ͻ15 Ͻ10 Ͻ10 ND R1249K Ͻ10 10 Ͻ15 Ͻ10 ND a The values shown are means of duplicate or triplicate determinations and are derived from Fig. 2, 4, and 5 (see figure legends for details).
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ABCC1 p.Arg1131Glu 15208328:118:478
status: NEW