ABCC1 p.Trp445Tyr
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PMID: 12388549
[PubMed]
Koike K et al: "Multiple membrane-associated tryptophan residues contribute to the transport activity and substrate specificity of the human multidrug resistance protein, MRP1."
No.
Sentence
Comment
48
Tryptophan substitutions were generated in the pGEM-3Z and pBluescriptSK(ϩ) plasmids above according to the manufacturer`s instructions with the following mutagenic primers (substituted nucleotides are underlined): MSD1 Trp mutants W40A (5Ј-G GTC CTC GTG GCC GTG CCT TG-3Ј, W47A (5Ј-GT TTT TAC CTC GCC GCC TGT TTC CCC-3Ј), W82A (5Ј-C TTG GGA TTT TTG CTG GCG ATC GTC TGC TGG GC-3Ј), W86A (5Ј-G CTG TGG ATC GTC TGC GCT GCA GAC CTC TTC TAC TC-3Ј), W94A (5Ј-C CTC TTC TAC TCT TTC GCG GAA AGA AGT CGG GGC-3Ј), W142A (5Ј-GGG ATC ATG CTC ACT TTC GCA CTG GTA GCC CTA ATG TG-3Ј), W142F (5Ј-G CTC ACT TTT TTC CTG GTA GCC C-3Ј); MSD2 Trp mutants W361A (5Ј-C ACG AAG GCC CCA GAT GCG CAG GGC TAC TTC TAC-3Ј), W445A (5Ј-G TAC ATT AAC ATG ATC GCG TCA GCC CCC CTG CAA G-3Ј), W445F (5Ј-CG TAC ATT AAC ATG ATC TTC TCA GCC CCC CTG CAA GTC-3Ј), W445Y (5Ј-CC ACG TAC ATT AAC ATG ATC TAC TCA GCG CCC CTG CAA GTC-3Ј), W459A (5Ј-GCT CTC TAC CTC CTG GCG CTG AAT CTG GGC CC-3Ј), W553A (5Ј-G GGC ACC TTC ACC GCG GTC TGC ACG CCC-3Ј), W553F (5Ј-G GGC ACC TTC ACC TTC GTC TGC ACG CCC-3Ј), W553Y (5Ј-GCC CTG GGC ACC TTC ACA TAT GTC TGC ACG CCC-3Ј; and MSD3 Trp mutants W1198A (5Ј-C GTG GCC AAC AGG GCG CTG GCC GTG CGG C-3Ј), W1198F (5Ј-GTG GCC AAC AGG TTC CTG GCC GTG CGG C-3Ј), W1198Y (5Ј-GTG GCC AAC AGG TAC CTG GCC GTG CGG C-3Ј).
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ABCC1 p.Trp445Tyr 12388549:48:951
status: NEW139 One mutant, W445Y, was reproducibly expressed at lower levels (approximately half) than the corresponding W445A and W445F mutants for reasons that are presently unclear.
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ABCC1 p.Trp445Tyr 12388549:139:12
status: NEW140 The most conservatively substituted MRP1-Trp445 mutant, W445Y, showed significant transport activity with respect to all five MRP1 substrates compared with the W445A mutant after correcting for differences in MRP1 expression levels.
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ABCC1 p.Trp445Tyr 12388549:140:56
status: NEW141 Thus, the LTC4 and E13SO4 transport activities of W445Y MRP1 were similar to those of wild-type MRP1, whereas transport of E217betaG, GSH, and MTX was 30-60% of wild-type activity (Fig. 6, B-F).
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ABCC1 p.Trp445Tyr 12388549:141:50
status: NEW142 The Phe-substituted Trp445 mutant, W445F, showed levels of transport intermediate between those of the W445A and W445Y mutants.
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ABCC1 p.Trp445Tyr 12388549:142:113
status: NEW189 Ala substitution of Trp1198 in MSD3 also resulted in a broad and profound decrease in MRP1 transport activity except for FIG. 6. ATP-dependent organic anion transport activity of wild-type and mutant MRP1 containing conservative Phe and Tyr substitutions of Trp1198 in TM16 of MSD3. A, immunoblot of membrane vesicles prepared from HEK293T cells transfected with empty vector (pcDNA3.1(-)), wild-type (WT-MRP1), and mutant (W445A, W445F, and W445Y; W553A, W553F and W553Y; W1198A, W1198F, and W1198Y) MRP1 cDNAs. MRP1 proteins were detected with monoclonal antibody QCRL-1, and relative levels of expression shown under the blot were estimated by densitometry.
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ABCC1 p.Trp445Tyr 12388549:189:442
status: NEW190 B-F, relative levels of 3 H-labeled organic anion uptake by the membrane vesicles shown in A enriched for wild-type MRP1 (solid bar), Trp 3 Phe-substituted MRP1 mutants (W445F, W553F, and W1198F; hatched bars), and Trp 3 Tyr-substituted MRP1 mutants (W445Y, W553Y, and W1198Y; open bars) were determined as described under "Experimental Procedures."
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ABCC1 p.Trp445Tyr 12388549:190:251
status: NEW