ABCMdb

ABCC1 p.Pro110Ala

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Publications

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

46 For the single substitutions of Pro with Ala in MSD1, the following sense mutagenic primers were used (substituted nucleotides are underlined): MRP1-P42A (5V-C GTG TGG GTG GCT TGT TTT TAC CTC TGG-3V), MRP1-P51A (5V-G GCC TGT TTC GCC TTC TAC TTC C-3V), MRP1-P69A (5V-CAG ATG ACA GCT CTC AAC-3V), MRP1-P104A (5V-C ATA TTC CTG GCC GCA GTG TTT CTG G-3V), MRP1-P110A (5V-GT TTC TGG TCA GCG CAA CTC TCT TGG-3V). Login to comment
104 Immunoblots of membrane vesicles prepared from the cell lines expressing MSD1 MRP1 mutants containing single (P42A, P51A, P69A and P110A) and double (P42/51A) Pro substitutions detected with MAb QCRL-1 are shown in Fig. 2A. The relative expression levels of the different mutants ranged from < 20% to approximately 150% that of wild-type MRP1. Login to comment
117 Similarly, the singly substituted MRP1-Pro mutants P42A, P51A, P69A, and P110A localized strongly to the plasma membrane, similar to wild-type MRP1 (MRP1-P42A and MRP1-P51A shown in Fig. 4A, middle two panels). Login to comment
180 Thus, all of the MSD1 Pro mutants retained the ability to transport LTC4 with TM1 mutants P42A, P51A, and P69A showing an approximately 30-50% decrease in relative LTC4 transport efficiency, while P110A (TM3) showed an increase of approximately 50%. Login to comment
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. Login to comment
193 E217hG uptake levels by the four single Pro MSD1 mutants P42A, P51A, P69A, and P110A were 48%, 43%, 95%, and 95% those of wild-type MRP1, respectively. Login to comment
179 Thus, all of the MSD1 Pro mutants retained the ability to transport LTC4 with TM1 mutants P42A, P51A, and P69A showing an approximately 30-50% decrease in relative LTC4 transport efficiency, while P110A (TM3) showed an increase of approximately 50%. Login to comment
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. Login to comment
192 E217hG uptake levels by the four single Pro MSD1 mutants P42A, P51A, P69A, and P110A were 48%, 43%, 95%, and 95% those of wild-type MRP1, respectively. Login to comment