ABCMdb

ABCC1 p.Asn19Gln

None has been submitted yet.

Publications

PMID: 16914551 [PubMed] Chen Q et al: "The amino terminus of the human multidrug resistance transporter ABCC1 has a U-shaped folding with a gating function."

No. Sentence Comment

196 It is also noteworthy that the mutants ABCC1FLAG-Q and ABCC1QQ , with the mutation of Asn19 to Gln, which is the last residue in the IU5C1 epitope, are still reactive to IU5C1 (Fig. 4, M and T). Login to comment

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."

No. Sentence Comment

66 The N19Q, N1006Q, and N1156Q mutations were generated using the TransformerTM site-directed mutagenesis kit (CLONTECH Laboratories, Inc., Palo Alto, CA) based on the method developed by Deng and Nickoloff (35). Login to comment
67 The templates for mutagenesis were prepared by cloning the BamHI fragment as above (for N19Q) and the XmaI fragment (MRP nucleotides 2337-4322) (for N1006Q and N1156Q) from pcDNAI-MRP1 into pGEM®-3Z (Promega, Madison, WI). Login to comment
68 Mutagenesis was then performed according to the manufacturer`s instructions using the ScaI/ StuI and SspI/EcoRV selection primers (for N19Q and N1006Q/N1156Q mutations, respectively), and the following sense mutagenic oligonucleotide primers: 5Ј-C TGG GAC TGG CAG GTC ACG TGG-3Ј (N19Q), 5Ј-C CCC ATC GTC CAG GGG ACT CAG G-3Ј (N1006Q), and 5Ј-C TAT TCC CAT TTC CAG GAG ACC TTG C-3Ј (N1156Q). Login to comment
69 The N19Q/N23Q double mutant was also generated by this method using the N19Q mutagenic primer with the N23Q BamHI fragment in pGEM¾1;-3Z as a template. Login to comment
72 The N19Q/N23Q/N1006Q triple mutant was prepared by cloning the BamHI fragment containing the N19Q/N23Q mutation into pcDNAI-MRP1/N1006Q. Login to comment
164 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). Login to comment
165 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). Login to comment
166 The MRP-(N19Q) and MRP-(N23Q) N-2 fragments are glycosylated because they have lower electrophoretic mobility than the deglycosylated 25-kDa N-2 fragment of wild-type MRP (compare with Fig. 4B, lane 1). Login to comment
168 To confirm that Asn19 and Asn23 are the only glycosylated sites in MSD1, we also produced an N19Q/N23Q MRP double mutant. Login to comment
169 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). Login to comment
178 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. Login to comment
70 The N19Q/N23Q double mutant was also generated by this method using the N19Q mutagenic primer with the N23Q BamHI fragment in pGEMt-3Z as a template. Login to comment