ABCC1 p.Arg1173Ala
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PMID: 19015228
[PubMed]
Conseil G et al: "Multiple roles of charged amino acids in cytoplasmic loop 7 for expression and function of the multidrug and organic anion transporter MRP1 (ABCC1)."
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Sentence
Comment
6
The properties of R1173A and E1184A were comparable with those of wild-type MRP1, whereas the remaining mutants were either poorly expressed (R1166A, D1183A) or exhibited reduced transport of one or more organic anions (E1144A, D1179A, K1181A, 1169 AAQA).
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ABCC1 p.Arg1173Ala 19015228:6:18
status: NEW59 Mutations were first 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 restriction site (substituted nucleotides are underlined): E1144A (5Ј-G AAG CGC CTC GCG TCG GTC AGC-3Ј); R1166A (5Ј C AGC GTC ATT GCA GCA TTC GAG GAG CAG-3Ј); R1166K (5Ј C AGC GTC ATT AAG GCC TTC GAG G-3Ј); 1169 EEQE-1169 AAQA (5Ј-C ATT CGA GCC TTC GCG GCA CAG GCA CGC TTC ATC C-3Ј); R1173A (5Ј-C GAG GAG CAG GAG GCA TTC ATC CAC CAG AG-3Ј); D1179A (5Ј-C CAC CAG AGT GCC CTT AAG GTG GAC G-3Ј), K1181A (5Ј-G AGT GAC CTG GCA GTC GAC GAG AAC C-3Ј); D1183A (5Ј-CTG AAG GTG GCC GAG AAC CAG-3Ј); D1183E (5Ј- CTG AAG GTG GAA GAG AAC CAG-3Ј); and E1184A (5Ј-GT GAC CTG AAG GTA GAC GCG AAC CAG AAG GCC-3Ј).
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ABCC1 p.Arg1173Ala 19015228:59:567
status: NEW158 For this reason, the R1173A and E1184A mutants were not investigated further.
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ABCC1 p.Arg1173Ala 19015228:158:21
status: NEW185 Shown is an immunoblot of membrane vesicles (1 g of protein) prepared from HEK 293T cells transfected with cDNA expression vectors encoding wild-type, E1144A, 1169 AAQA, R1173A, D1179A, K1181A, and E1184A MRP1 mutants.
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ABCC1 p.Arg1173Ala 19015228:185:178
status: NEW191 Of the seven single-Ala-substituted CL7 mutants generated, only two (R1173A and E1184A) displayed properties that were comparable with those of wild-type MRP1, demonstrating the extreme sensitivity of this cytoplasmic loop to mutation.
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ABCC1 p.Arg1173Ala 19015228:191:69
status: NEW212 Percentage Wild-Type MRP1 Uptake Activity E1144A 1169 AAQA R1173A D1179A K1181A E1184A % LTC4 90 Ϯ 5 70 Ϯ 5 120 Ϯ 5 65 Ϯ 5 55 Ϯ 5 85 Ϯ 5 E217bG 55 Ϯ 5 80 Ϯ 0 130 Ϯ 10 90 Ϯ 5 65 Ϯ 5 95 Ϯ 15 E13SO4 45 Ϯ 5 45 Ϯ 5 125 Ϯ 0 65 Ϯ 0 50 Ϯ 5 90 Ϯ 10 MTX 55 Ϯ 5 105 Ϯ 5 N.D. N.D. 45 Ϯ 5 N.D. GSH 80 Ϯ 5 95 Ϯ 10 N.D. N.D. 30 Ϯ 5 N.D. N.D., not determined.
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ABCC1 p.Arg1173Ala 19015228:212:59
status: NEW