ABCC1 p.Arg1222Met
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PMID: 12450376
[PubMed]
Ren XQ et al: "A positively charged amino acid proximal to the C-terminus of TM17 of MRP1 is indispensable for GSH-dependent binding of substrates and for transport of LTC4."
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72
MRP1 constructs encoding R1222M and R1249A were generated in a PCR using the forward primers 5'-CATG- CACAGCCTCAGTGCTG-3' and 5'-GCGATGTCATCT- GAAATGGAAACC-3', respectively (bold denotes mismatched bases encoding the mutations).
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ABCC1 p.Arg1222Met 12450376:72:25
status: NEW73 An EcoRV site (underlined) was created by silent mutation (bold) in the R1222M construct using the reverse primer 5'-GATATC- ACCGCAAACAGGGCAGC-3'.
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ABCC1 p.Arg1222Met 12450376:73:72
status: NEW78 The EcoRV-KpnI fragment from the construct encoding R1222M was cloned into the multiple cloning site II of the pFastBac DUAL plasmid between the SmaI and KpnI sites.
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ABCC1 p.Arg1222Met 12450376:78:52
status: NEW169 Arginine at position 1222 in the azido AG-A-photolabeled MRP11-1222 fragment was replaced with Met, and the functions of MRP1 R1222M were compared with those of MRP1 R1249A.
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ABCC1 p.Arg1222Met 12450376:169:126
status: NEW190 Figure 6A shows that the expression level of MRP1 R1222M was comparable to that of wild-type MRP1, whereas the expression level of MRP1 R1249A was higher than that of wild-type MRP1.
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ABCC1 p.Arg1222Met 12450376:190:50
status: NEW192 As shown in Figure 6B, MRP1 R1222M transported LTC4 as efficiently as wild-type MRP1.
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ABCC1 p.Arg1222Met 12450376:192:28
status: NEW195 As shown in Figure 6C, GSH stimulated [125 I]azido AG-A binding to MRP1 R1222M as efficiently as wild-type MRP1.
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ABCC1 p.Arg1222Met 12450376:195:72
status: NEW225 Membrane vesicles (25 µg of protein) expressing MRP1 R1222M or MRP1 R1249A as indicated were incubated with 1.37 nM [3H]LTC4 at 37 °C in 50 µL of transport buffer as described in the legend of Figure 3 in the presence or absence of 4 mM ATP at the indicated periods.
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ABCC1 p.Arg1222Met 12450376:225:58
status: NEW229 Membrane vesicles (100 µg of protein) expressing MRP1, MRP1 R1249M, and MRP1 R1222M were photolabeled with [125I]azido AG-A in the absence or presence of the indicated concentrations of GSH as described in the legend ofo Figure 2.
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ABCC1 p.Arg1222Met 12450376:229:82
status: NEW