ABCC1 p.Pro272Ala

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PMID: 12948592 [PubMed] Ito K et al: "Mutation of proline residues in the NH(2)-terminal region of the multidrug resistance protein, MRP1 (ABCC1): effects on protein expression, membrane localization, and transport function."
No. Sentence Comment
47 For substitution of Pro residues in the CL3 loop connecting MSD1 to MSD2, the mutagenic primers were: MRP1-P196A (5V-CA GAT CGC TCA GCC CTG TTC TC-3V), MRP1-P205A (5V-C ATC CAC GAC GCT AAT CCC TG-3V), MRP1-P207A (5V-C GAC CCT AAT GCC TGC CCA GAG-3V), MRP1-P209A (5V-CTA ATC CCT GCG CAG AGT CCA G-3V), MRP1-P235A (5V-C TAC CGC CAG GCC CTG GAG GG-3V), MRP1-P255A (5V-CG GAA CAA GTC GTG GCT GTT TTG G-3V), MRP1-P272A (5V-CT AGG AAG CAG GCG GTG AAG G-3V).
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ABCC1 p.Pro272Ala 12948592:47:408
status: NEW
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ABCC1 p.Pro272Ala 12948592:47:420
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107 The seven single (P196A, P205A, P207A, P209A, P235A, P255A, and P272A) and two multiply substituted CL3 mutants (P196/205/207/209A and P235/255A) could also be stably expressed in HeLa cells.
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ABCC1 p.Pro272Ala 12948592:107:64
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149 Duplicate immunoblots of the P235A, P255A, P235/255A, and P272A MRP1 mutants probed with MAb QCRL-1 and MAb MRPr1 indicated that detection of the P255A and P235/P255A mutant MRP1 proteins relative to wild-type MRP1 by MAb MRPr1 was lower than that by MAb QCRL-1 (not shown).
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ABCC1 p.Pro272Ala 12948592:149:58
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188 Results were expressed relative to wild-type Table 1 Kinetic parameters of [3 H]LTC4 uptake by membrane vesicles prepared from transfected HeLa cells expressing MRP1-Pro mutants Transfected cell line Km (nM) Vmax (pmol minÀ 1 mgÀ 1 ) Normalized Vmax MSD1 mutants WT-MRP1 128 80 80 P42A 139 35 41 P51A 109 55 50 P42/51A 103 15 48 P69A 142 46 42 P110A 150 73 124 CL3 mutants WT-MRP1 88 57 57 P196A 143 25 52 P205A 63 14 88 P207A 81 75 110 P209A 112 121 85 P196/205/207/209A 90 87 94 P235A 91 45 30 P255A 99 110 67 P235/255A 97 81 35 P272A 80 59 109 The kinetic parameters of [3 H]LTC4 uptake were determined by regression analysis of the Eadie-Hofstee transformed data.
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ABCC1 p.Pro272Ala 12948592:188:541
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148 Duplicate immunoblots of the P235A, P255A, P235/255A, and P272A MRP1 mutants probed with MAb QCRL-1 and MAb MRPr1 indicated that detection of the P255A and P235/P255A mutant MRP1 proteins relative to wild-type MRP1 by MAb MRPr1 was lower than that by MAb QCRL-1 (not shown).
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ABCC1 p.Pro272Ala 12948592:148:58
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187 Results were expressed relative to wild-type Table 1 Kinetic parameters of [3 H]LTC4 uptake by membrane vesicles prepared from transfected HeLa cells expressing MRP1-Pro mutants Transfected cell line Km (nM) Vmax (pmol min 1 mg 1 ) Normalized Vmax MSD1 mutants WT-MRP1 128 80 80 P42A 139 35 41 P51A 109 55 50 P42/51A 103 15 48 P69A 142 46 42 P110A 150 73 124 CL3 mutants WT-MRP1 88 57 57 P196A 143 25 52 P205A 63 14 88 P207A 81 75 110 P209A 112 121 85 P196/205/207/209A 90 87 94 P235A 91 45 30 P255A 99 110 67 P235/255A 97 81 35 P272A 80 59 109 The kinetic parameters of [3 H]LTC4 uptake were determined by regression analysis of the Eadie-Hofstee transformed data.
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ABCC1 p.Pro272Ala 12948592:187:529
status: NEW
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