ABCC1 p.Asn71Gln
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PMID: 9295302
[PubMed]
Hipfner DR et al: "Membrane topology of the multidrug resistance protein (MRP). A study of glycosylation-site mutants reveals an extracytosolic NH2 terminus."
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Sentence
Comment
61
The N23Q and N71Q mutations were generated by a modification of the PCR-based megaprimer method (34).
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ABCC1 p.Asn71Gln 9295302:61:13
status: NEW63 The first PCR was carried out on the pBluescript construct containing the BamHI fragment cloned in the EcoRI to HindIII orientation (5Ј 3 3Ј) using the M13 primer and either the N23Q (5Ј-C ACG TGG CAA ACC AGC AAC CCC GAC T-3Ј) or the N71Q (5Ј-A CCT CTC CAG AAA ACC AAA ACT GCC T-3Ј) mutagenic primer (substituted nucleotide positions underlined).
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ABCC1 p.Asn71Gln 9295302:63:258
status: NEW164 Introduction of the individual N19Q, N23Q, and N71Q mutations in MSD1 did not result in a detectable alteration of the electrophoretic mobility of the intact proteins (Fig. 4A, lanes 3-5).
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ABCC1 p.Asn71Gln 9295302:164:47
status: NEW165 However, after digestion with trypsin, the N-2 fragments of wild-type MRP and MRP-(N71Q) co-migrated at 43-60 kDa, whereas the N-2 fragments of MRP-(N19Q) and MRP-(N23Q) migrated slightly faster at approximately 38-50 kDa (Fig. 4B, lanes 2-5).
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ABCC1 p.Asn71Gln 9295302:165:83
status: NEW169 MRP-(N19Q/ N23Q) migrated faster than wild-type MRP and the N19Q, N23Q, and N71Q single mutants (Fig. 4A, compare lane 6 with lanes 2-5).
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ABCC1 p.Asn71Gln 9295302:169:76
status: NEW178 A, crude membrane proteins from Cos-1 cells expressing wild-type MRP (WT) treated with (ϩ) or without (-) PNGase F, the single glycosylation site mutant MRP constructs N19Q, N23Q, N71Q, N1006Q, and N1156Q, the double mutant N19Q/N23Q, and the triple mutant N19Q/N23Q/N1006Q were separated by SDS-polyacrylamide gel electrophoresis on 5% gels and immunoblotted with mAb QCRL-1.
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ABCC1 p.Asn71Gln 9295302:178:180
status: NEWX
ABCC1 p.Asn71Gln 9295302:178:186
status: NEW62 The N23Q and N71Q mutations were generated by a modification of the PCR-based megaprimer method (34).
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ABCC1 p.Asn71Gln 9295302:62:13
status: NEW64 The first PCR was carried out on the pBluescript construct containing the BamHI fragment cloned in the EcoRI to HindIII orientation (59 3 39) using the M13 primer and either the N23Q (59-C ACG TGG CAA ACC AGC AAC CCC GAC T-39) or the N71Q (59-A CCT CTC CAG AAA ACC AAA ACT GCC T-39) mutagenic primer (substituted nucleotide positions underlined).
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ABCC1 p.Asn71Gln 9295302:64:234
status: NEW