ABCC1 p.Asp1183Ala
[switch to full view]Comments [show]
None has been submitted yet.
PMID: 19015228
[PubMed]
Conseil G et al: "Multiple roles of charged amino acids in cytoplasmic loop 7 for expression and function of the multidrug and organic anion transporter MRP1 (ABCC1)."
No.
Sentence
Comment
6
The properties of R1173A and E1184A were comparable with those of wild-type MRP1, whereas the remaining mutants were either poorly expressed (R1166A, D1183A) or exhibited reduced transport of one or more organic anions (E1144A, D1179A, K1181A, 1169 AAQA).
X
ABCC1 p.Asp1183Ala 19015228:6:150
status: NEW59 Mutations were first generated in the pGEM-3Z-XmaI/MRP1 plasmid according to the manufacturer`s instructions with the following mutagenic primers (Integrated DNA Technologies, Inc., Coralville, IA), which also introduced an additional restriction site (substituted nucleotides are underlined): E1144A (5Ј-G AAG CGC CTC GCG TCG GTC AGC-3Ј); R1166A (5Ј C AGC GTC ATT GCA GCA TTC GAG GAG CAG-3Ј); R1166K (5Ј C AGC GTC ATT AAG GCC TTC GAG G-3Ј); 1169 EEQE-1169 AAQA (5Ј-C ATT CGA GCC TTC GCG GCA CAG GCA CGC TTC ATC C-3Ј); R1173A (5Ј-C GAG GAG CAG GAG GCA TTC ATC CAC CAG AG-3Ј); D1179A (5Ј-C CAC CAG AGT GCC CTT AAG GTG GAC G-3Ј), K1181A (5Ј-G AGT GAC CTG GCA GTC GAC GAG AAC C-3Ј); D1183A (5Ј-CTG AAG GTG GCC GAG AAC CAG-3Ј); D1183E (5Ј- CTG AAG GTG GAA GAG AAC CAG-3Ј); and E1184A (5Ј-GT GAC CTG AAG GTA GAC GCG AAC CAG AAG GCC-3Ј).
X
ABCC1 p.Asp1183Ala 19015228:59:764
status: NEW120 These experiments showed that two of the Ala-substituted mutants (R1166A and D1183A) were consistently expressed at very low levels (10-20% of wild-type MRP1) (Fig. 2A).
X
ABCC1 p.Asp1183Ala 19015228:120:77
status: NEW124 Confocal microscopy of intact HEK cells using the MRP1-specific mAb QCRL-3 showed that despite their low expression levels, at least a portion of the R1166A and D1183A mutant proteins were correctly routed to the plasma mem- brane (not shown).
X
ABCC1 p.Asp1183Ala 19015228:124:161
status: NEW126 However, even at the lower temperature, expression of the R1166A and D1183A mutants remained substantially below that of wild-type MRP1; furthermore, plasma membrane routing of the mutants as well as wild-type MRP1 was impaired (data not shown).
X
ABCC1 p.Asp1183Ala 19015228:126:69
status: NEW138 A, immunoblot of whole cell lysates (10 g of protein) prepared from HEK 293T cells transfected with wild-type (WT-MRP1), and R1166A and D1183A mutant MRP1 expression vectors.
X
ABCC1 p.Asp1183Ala 19015228:138:144
status: NEW140 MRP1 levels in whole-cell lysates were detected with mAb QCRL-1, and the relative protein expression levels are shown under the blot and were estimated by densitometry as described under Materials and Methods. B, immunoblots of whole-cell lysates (10 g of protein) prepared from HEK 293T cells transfected with R1166A/K and D1183A/E mutant and wild-type (WT) MRP1 cDNA expression vectors. MRP1 proteins were detected with mAb QCRL-1 and lysates of untransfected cells were included as a negative control as above.
X
ABCC1 p.Asp1183Ala 19015228:140:332
status: NEW192 The remaining five single mutants were either poorly expressed (R1166A, D1183A) (Fig. 2) or exhibited significantly reduced transport levels of one or more organic anions (E1144A, D1179A, K1181A) (Table 1).
X
ABCC1 p.Asp1183Ala 19015228:192:72
status: NEW194 The poor expression of the R1166A and D1183A mutants indicates that Arg1166 and Asp1183 contribute to the stability of MRP1, probably by influencing the proper folding of the transporter during its biosynthesis.
X
ABCC1 p.Asp1183Ala 19015228:194:38
status: NEW199 Elucidation of the post-translational events leading to poor expression of R1166A and D1183A and other low or nonexpressing MRP1 mutants is currently under investigation.
X
ABCC1 p.Asp1183Ala 19015228:199:86
status: NEW201 Our observation that the same-charge D1183E mutant was as poorly expressed at the plasma membrane as the neutrally substituted D1183A is somewhat unexpected, because previously identified expression impaired mutants involving charged residues have typically not exhibited this property (Haimeur et al., 2004; Situ et al., 2004).
X
ABCC1 p.Asp1183Ala 19015228:201:127
status: NEW244 Unlike the R1166A and D1183A mutants, the second group of functionally altered single Ala-substituted mutants (E1144A, D1179A, K1181A) and the 1169 AAQA triple mutant were all expressed at levels comparable with or greater than that of wild-type MRP1 (Fig. 4).
X
ABCC1 p.Asp1183Ala 19015228:244:22
status: NEW
PMID: 21143116
[PubMed]
He SM et al: "Structural and functional properties of human multidrug resistance protein 1 (MRP1/ABCC1)."
No.
Sentence
Comment
809
The Arg1166Ala and Asp1183Ala/Glu mutants were poorly expressed, probably by affecting the proper folding of the protein during its biosynthesis.
X
ABCC1 p.Asp1183Ala 21143116:809:19
status: NEW