ABCMdb

ABCC1 p.Asn1156Gln

None has been submitted yet.

Publications

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 pGEM௡-3Z as a template. Login to comment
172 We next examined expression of the MSD3 glycosylation acceptor site mutants MRP-(N1006Q) and MRP-(N1156Q). Login to comment
173 The MRP-(N1156Q) mutant co-migrated with the wild-type protein, whereas MRP-(N1006Q) displayed a greater electrophoretic mobility (Fig. 4A, lanes 2, 7 and 8). Login to comment
174 The C-1 fragments derived from wild-type MRP and MRP-(N1156Q) also co-migrated, and the C-1 fragment of MRP-(N1006Q) had the same electrophoretic mobility as the PNGaseF-treated wild-type C-1 fragment (Fig. 4C, lanes 1-4). 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
231 On the other hand, it was expected that the MRP-(N1156Q) mutant might be informative because this asparagine residue is predicted to be extracytoplasmic only in the MSD3 configurations with four transmembrane segments. Login to comment