ABCMdb

ABCC2 p.Trp1254Tyr

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Publications

PMID: 11925441 [PubMed] Zhang DW et al: "Determinants of the substrate specificity of multidrug resistance protein 1: role of amino acid residues with hydrogen bonding potential in predicted transmembrane helix 17."

No. Sentence Comment

191 However, when the corresponding residue, Trp1254 , is mutated in MRP2, only nonconserved substitutions eliminate E217betaG transport, and in contrast to MRP1, only the most conservatively substituted MRP2 Trp1254 mutant, W1254Y, transports LTC4 (45). Login to comment

PMID: 14965249 [PubMed] Haimeur A et al: "The MRP-related and BCRP/ABCG2 multidrug resistance proteins: biology, substrate specificity and regulation."

No. Sentence Comment

395 Furthermore, only the most conservatively substituted Trp1254 mutant (MRP2-Trp1254Tyr) retained any LTC4 transport activity whereas transport of this substrate by the comparable MRP1-Trp1246 mutants was similar to wild-type MRP1. Login to comment

PMID: 22290738 [PubMed] Arlanov R et al: "Functional characterization of protein variants of the human multidrug transporter ABCC2 by a novel targeted expression system in fibrosarcoma cells."

No. Sentence Comment

277 Moreover, a substitution of W1254 by conserved and nonconserved amino acids (e.g., W1254A, W1254Y) in TM17 seems to be critical for substrate binding and transport activity [Ito et al., 2001b]. Login to comment

PMID: 20082599 [PubMed] Jemnitz K et al: "ABCC2/Abcc2: a multispecific transporter with dominant excretory functions."

No. Sentence Comment

97 Mutant Predicted location Substrate Activity changes Reference Human MRP2 Δ1-188 TMD0 LTC4 ↓ Fernandez et al., 2002 K316A JC, TM6 GMF ↔ Ryu et al., 2000 K324A TM6 GMF ↓ Ryu et al., 2000 K329A TM6 GMF ↔ Ryu et al., 2000 R412G DJ IC MTX ↓ Hulot et al., 2005 W417I IC, TM7-TM8 E2-17βG ↓ Hirouchi et al., 2004 LTC4 ↓ DNP-SG ↓ H439A TM8 GMF ↔ Ryu et al., 2000 K483A IC, JM, TM9 GMF ↓ Ryu et al., 2000 K590A JC, TM11 GMF ↔ Ryu et al., 2000 S789F NBD1 E2-17βG ↓ Hirouchi et al., 2004 LTC4 ↓ DNP-SG ↓↓ R1023A EC, JM, TM13 GMF ↔ Ryu et al., 2000 H1042A TM13 GMF ↔ Ryu et al., 2000 R1100A JC, TM14 GMF ↔ Ryu et al., 2000 P1158A IC, JM, TM15 LTC4 ↓↓ Letourneau et al., 2007 E2-17βG ↔ MTX ↔ Table 1. continued on next page Mutant Predicted location Substrate Activity changes Reference I1173F DJ IC, TM15-16 LTC4 No act Keitel et al., 2003 E2-17βG No act R1210A EC, JC, TM16 GMF ↓↓ Ryu et al., 2000 R1230A TM16 GMF ↔ R1257A JC, TM17 GMF ↓↓ W1254A JC, TM17 E2-17βG ↓↓ Ito et al., 2001a W1254C ↓↓ W1254F ↔ W1254Y ↔ W1254A JC, TM17 LTC4 ↓↓ Ito et al., 2001b W1254C ↓↓↓ W1254F ↓↓ W1254Y ↓↓ W1254A JC, TM17 MTX ↓↓ Ito et al., 2001a W1254C ↓↓ W1254F ↓↓ W1254Y ↓↓↓ A1450T NBD2 E2-17βG ↓↓ Hirouchi et al., 2004 LTC4 ↓↓ DNP-SG ↓↓ Rat Mrp2 K308M IC, JM, TM6 TLC-S ↔ Ito et al., 2001b DNP-G ↑ LTC4 ↓ E3040G ↔ K320M TM6 TLC-S ↑ DNP-G ↑ LTC4 ↓ E3040G ↑ K325M TM6 TLC-S ↓* DNP-G ↓↓↓* LTC4 ↓↓↓* E3040G ↓ D329N TM6 TLC-S ↔ DNP-G ↓ LTC4 ↓↓↓* E3040G ↓ R586L TM11 TLC-S ↓ DNP-G ↓↓* LTC4 ↓↓* E3040G ↔ R1019M IC, JM, TM13 TLC-S ↔ DNP-G ↑* LTC4 ↔ E3040G ↔ R1096L TM14 TLC-S ↑ DNP-G ↑ LTC4 ↔ E3040G ↔ EC, extracellular; IC, intracellular; JC, near the cytosol in the membrane; JM, juxtamembrane; TLC-S, tauro-litocholate-sulfate; GMF, glutathione- methyl-fluorescein; ↑, activity over control>1.2; ↔, 1.2>activity over control>0.8; ↓, 0.8>activity over control>0.5; ↓↓, 0.5>activity over control>0.1; ↓↓↓, 0.1>activity over control. Login to comment

PMID: 12406647 [PubMed] Suzuki H et al: "Single nucleotide polymorphisms in multidrug resistance associated protein 2 (MRP2/ABCC2): its impact on drug disposition."

No. Sentence Comment

126 It was found that Lys325Met and ly substituted Trp1254Tyr transports E 17bG and2 Arg586Leu resulted in a marked reduction in the LTC , but not methotrexate (Fig. 2) [112].4 transport of glutathione-conjugates, whereas the In addition to the previously described mutation transport activity for glucuronide conjugates was found in a DJS patient, the carboxy-terminal se- retained (Fig. 2) [109]. Login to comment

PMID: 12130697 [PubMed] Gerk PM et al: "Regulation of expression of the multidrug resistance-associated protein 2 (MRP2) and its role in drug disposition."

No. Sentence Comment

57 Only the most conservative substitution (W1254Y) retained leukotriene C4 transport activity. Login to comment

PMID: 11500505 [PubMed] Ito K et al: "Mutation of Trp1254 in the multispecific organic anion transporter, multidrug resistance protein 2 (MRP2) (ABCC2), alters substrate specificity and results in loss of methotrexate transport activity."

No. Sentence Comment

6 In addition, only the most conservatively substituted mutant (W1254Y) transported [3 H]leukotriene C4, whereas all other substitutions eliminated transport of this substrate. Login to comment
47 Mutagenesis was carried out using the TransformerTM kit (CLONTECH, Palo Alto, CA) according to the manufacturer`s instructions with the following sense mutagenic primers (substituted nucleotides are underlined): W1254A (5Ј-C- ACAAACCCTGAACGCTTCTGGTGAGGATGAC-3Ј), W1254C (5Ј-CAC- AAACCCTGAACTGTCTGGTGAGGATGAC-3Ј), W1254F (5Ј-CACAAA- CCCTGAACTTTCTGGTGAGGATGAC-3Ј), and W1254Y (5Ј-CACAAA- CCCTGAACTATCTGGTGAGGATGAC-3Ј). Login to comment
74 These included nonconservative substitutions with a nonaromatic nonpolar amino acid (Ala; W1254A-MRP2) and a nonaromatic polar amino acid (Cys; W1254C-MRP2), as well as conservative substitutions with polar (Tyr; W1254Y-MRP2) and nonpolar (Phe; W1254F-MRP2) aromatic amino acids. Login to comment
76 As shown in Fig. 1B, wild-type MRP2 and its four mutants (W1254A, W1254C, W1254Y, and W1254F) were all expressed in the HEK293T cells at comparable levels. Login to comment
77 Mean values of expression levels from four to six independent transfections relative to wild-type MRP2 were W1254A, 1.1; W1254C, 0.9; W1254F, 0.9; W1254Y, 1.0, respectively (Fig. 1B). Login to comment
81 However, E217betaG transport by the W1254Y-MRP2 and W1254F-MRP2 mutants was similar to that of wild-type MRP2, in marked contrast to the comparable conservatively substituted MRP1-Trp1246 mutants, which were inactive for transport of this substrate (40). Login to comment
83 The [3 H]LTC4 transport activity of the fourth and most conservatively substituted mutant, W1254Y-MRP2, was ϳ40% of wild-type MRP2. Login to comment
97 B, immunoblot of membrane vesicles prepared from HEK293T cells transfected with empty vector (pcDNA3.1(-)), wild-type (WT-MRP2) and mutant (W1254A, W1254C, W1254F, and W1254Y) MRP2 cDNAs. Login to comment
103 A, [3 H]E217betaG uptake was measured in membrane vesicles prepared from HEK293T cells transfected with empty vector (pcDNA3.1(-); open bar), wild-type MRP2 (WT-MRP2; solid bar), and mutant (W1254A-MRP2, W1254C-MRP2, W1254F-MRP2, and W1254Y-MRP2; shaded bars) cDNA expression vectors. Login to comment
106 The results shown are the means Ϯ S.D. of triplicate determinations in a single experiment, and similar results were obtained in a second experiment. relative levels of [3 H]MTX transport by the W1254A, W1254C, W1254F, and W1254Y mutants were 14, 14, 11, and 1%, respectively, of wild-type MRP2. Login to comment
111 To determine whether E217betaG transport by mutant MRP2 proteins that no longer transport MTX remained inhibitable by this antifolate, [3 H]E217betaG transport in membrane vesicles prepared from cells expressing W1254F-MRP2 and W1254Y-MRP2 was examined. Login to comment
112 In contrast to wild-type MRP2, MTX had no significant effect on [3 H]E217betaG transport by either W1254F-MRP2 or W1254Y-MRP2 at concentrations of either 500 or 1000 ␮M MTX (Fig. 4B). Login to comment
116 In addition, we observed that [3 H]E217betaG transport by the MRP2 mutants W1254A, W1254C, W1254F, and W1254Y was enhanced 9-, 6-, 9-, and 10-fold, respectively, by sulfinpyrazone compared with 4-fold for wild-type MRP2. Login to comment
133 B, membrane vesicles (8 ␮g of protein) from cells expressing WT-MRP2 and mutants W1254F-MRP2 and W1254Y-MRP2 were incubated at 37 °C with 400 nM [3 H]E217betaG in transport buffer and other components for 5 min in the absence (open bars) or the presence of MTX (500 ␮M, shaded bar; 1000 ␮M, solid bar) as described under "Experimental Procedures." Login to comment
151 However, although [3 H]E217betaG transport by the MRP2- W1254F and MRP2-W1254Y mutants appeared unchanged despite their inability to transport MTX, transport of the conjugated estrogen was no longer inhibited by the antifolate agent. Login to comment
156 Of the three MRP2 substrates examined in the present study, none were transported by the nonconservatively substituted W1254C and W1254A mutants, one was transported by the W1254F mutant, and two were transported by the W1254Y mutant. Login to comment
159 Effect of sulfinpyrazone on [3 H]E217betaG uptake and [3 H]MTX uptake by wild-type and Trp1254 mutant MRP2 proteins. A, [3 H]E217betaG uptake was measured in the absence (open bars) and the presence (solid and shaded bars) of sulfinpyrazone (100 ␮M) in membrane vesicles prepared from cells expressing wild-type (WT-MRP2) and mutant (W1254A, W1254C, W1254F, and W1254Y) MRP2 proteins as described under "Experimental Procedures." Login to comment