ABCMdb

ABCC1 p.Glu535Asp

None has been submitted yet.

Publications

PMID: 21177244 [PubMed] Iram SH et al: "Expression and function of human MRP1 (ABCC1) is dependent on amino acids in cytoplasmic loop 5 and its interface with nucleotide binding domain 2."

No. Sentence Comment

47 To generate point mutations in CL5, plasmid pBluescriptSK(ϩ)BamHI/SpHI-MRP1 containing a 1.9-kb fragment from pcDNA3.1(-)-MRP1k was used as the template with the following mutagenic primers (substituted nucleotides are underlined): R501A, 5Ј-GAG CAA AGA CAA TGC GAT CAA GCT GAT G-3Ј; K503A, 5Ј-GAC AAT CGG ATC GCG CTG ATG AAC G-3Ј; E507A, 5Ј-GCT GAT GAA CGC AAT TCT CAA TGG G-3Ј; G511I, 5Ј-CTC AAT ATT ATC AAA GTG CTA AAG CTT TAT GCC TGG GAG CTG GC-3Ј; K513A, 5Ј-CTC AAT GGG ATC GCA GTG CTA AAG C-3Ј; K513R, 5Ј-CTC AAT GGG ATC AGA GTG CTA AAG C-3Ј; K516A, 5Ј-GAT CAA AGT GCT AGC GCT TTA TGC CTG-3Ј; K516R, 5Ј-GAT CAA AGT GCT AAG ACT TTA TGC CTG-3Ј; E521A, 5Ј-CTT TAT GCC TGG GCG CTG GCA TTC-3Ј; E521D, 5Ј-CTT TAT GCC TGG GAC CTG GCA TTC-3Ј; R532A, 5Ј-GTG CTG GCC ATT GCG CAG GAG GAG CT-3Ј; E535A, 5Ј-CAT CAG GCA GGA GGC CTT GAA GGT GCT GAA G-3Ј; and E535D, 5Ј- CAG GCA GGA GGA TCT GAA GGT GCT GAA G-3Ј. Login to comment
141 Expression and Transport Activity of "Same Charge" Mutants of Lys513 , Lys516 , Glu521 , and Glu535 -To determine whether it was the charge or another property of the amino acid at positions 513, 516, 521, and 535 that was critical for efficient plasma membrane expression of MRP1, same charge mutants (K513R, K516R, E521D, and E535D) were created and again expressed in HEK cells. Login to comment
142 At 37 °C, levels of the K513R mutant were comparable with wild-type MRP1 levels, whereas levels of the K516R, E521D, and E535D mutants were somewhat reduced (20-40%) (Fig. 4A). Login to comment
148 A and B, representative immunoblots of whole cell lysates (WCL) (10 ␮g of protein/lane) (A) and membrane vesicles (1 ␮g of protein/lane) (B) prepared from HEK293T cells transfected with wild-type (WT-MRP1) and mutant (K513R, K516R, E521D, and E535D) MRP1 cDNA expression vectors. Untransfected cells were used as a negative control. MRP1 levels were detected with mAb QCRL-1, and the relative protein expression levels, estimated by densitometry, are shown below the blots. Login to comment