ABCMdb

ABCC1 p.Cys208Ala

None has been submitted yet.

Publications

PMID: 12731862 [PubMed] Leslie EM et al: "Functional and structural consequences of cysteine substitutions in the NH2 proximal region of the human multidrug resistance protein 1 (MRP1/ABCC1)."

No. Sentence Comment

110 Those exhibiting minor disruption of plasma membrane trafficking included cell lines expressing four CysfAla mutants [Cys43Ala (Figure 3C), Cys85Ala (Figure 3G), Cys190Ala (Figure 3K), and Cys208Ala (Figure 3M)] and four CysfSer mutants [Cys49Ser (Figure 3F), Cys85Ser (Figure 3H), Cys190Ser (Figure 3L), and Cys208Ser (Figure 3N)]. Login to comment
130 Table 1: Detection of Tryptic Fragments N1 and N2 of MRP1 in Membranes Prepared from HeLa Cells Stably Expressing CysfAla and CysfSer MRP1 Mutantsa trypsin:protein ratio (w:w)transfected HeLa cell line N1 detected N2 detected WT-MRP1 1:10 000 1:1000 C43A-MRP1 1:100 1:100 C43S-MRP1 1:10 000 1:1000 C49A-MRP1 1:250 1:100 C49S-MRP1 1:10 000 1:500 C85A-MRP1 1:10 000 1:1000 C85S-MRP1 1:1000 1:250 C148A-MRP1 1:250 1:250 C148S-MRP1 1:1000 1:500 C190A-MRP1 1:1000 1:1000 C190S-MRP1 1:1000 1:250 C208A-MRP1 1:10 000 1:250 C208S-MRP1 1:10 000 1:500 C265A-MRP1 1:250 1:10 C265S-MRP1 1:1000 1:250 a The data shown represent a summary of the limited trypsin digests shown in Figures 4 and 5. Login to comment
132 Cys49Ser, Cys190Ala, Cys208Ala, and Cys208Ser also closely matched those of WT-MRP1 except that the N1 fragment did not appear until a trypsin:protein ratio of 1:1000 for Cys190Ala-MRP1 and the N2 fragments of Cys49Ser-MRP1, Cys208Ala-MRP1, and Cys208Ser-MRP1 did not appear until trypsin:protein ratios of 1:500, 1:250, and 1:500, respectively. The remaining CysfSer MRP1 mutants (Cys85Ser, Cys148Ser, Cys190Ser, and Cys265Ser) were more resistant to trypsinolysis than WT-MRP1 but less resistant than their respective Ala partners. Login to comment
147 Ala substitution of Cys49 resulted in a 37% decrease in LTC4 transport activity while replacement of Cys208 and Cys265 with Ala resulted in an approximate 30% increase in activity. Four of the seven CysfSer MRP1 mutants also transported LTC4 at levels comparable to those of WT-MRP1; however, some differences between the transport activities of the Ala and Ser substituted mutants were noted (Figure 6B). Login to comment
150 Similar to Cys208Ala-MRP1, Cys208Ser-MRP1 LTC4 transport activity was increased 30%. Login to comment
158 In addition, unlike Cys208Ala-MRP1, which transported E217 G at a level similar to WT-MRP1, E217 G uptake by Cys208Ser-MRP1 was reduced by 35%. Login to comment
173 Consequently, we examined the resistance of cells expressing mutant MRP1 molecules harboring substitutions of Cys residues in MSD1 (Cys43Ala, Cys43Ser, Cys49Ala, Cys49Ser, Cys190Ala, and Cys190Ser) andCL3(Cys208Ala,Cys208Ser,Cys265Ala,andCys265Ser), to sodium arsenite and potassium antimony tartrate. Login to comment
192 Table 2: Sensitivity of Stably Transfected HeLa Cells Expressing Wild-Type and Cys-Substituted MRP1 to Sodium Arsenite and Potassium Antimony Tartrate relative resistancea transfected HeLa cell line Na+ arsenite K+ antimony tartrate WT-MRP1 3.6 ( 1.3 (1) (n ) 6) 2.0 ( 0.5 (1) (n ) 7) C43A-MRP1 4.3 ( 0.7 (1.2) (n ) 3) 2.0 ( 0.4 (1) (n ) 3) C43S-MRP1 1.4 ( 0.5 (0.4)b (n ) 6) 2.8 ( 1.1 (1.4) (n ) 4) C49A-MRP1 4.0 ( 1.6 (1.1) (n ) 5) 2.6 ( 0.7 (1.3) (n ) 4) C49S-MRP1 3.0 ( 1.5 (0.8) (n ) 4) 3.0 ( 0.6 (1.5) (n ) 4) C190A-MRP1 3.8 ( 0.4 (1) (n ) 4) 4.0 ( 1.2 (2) (n ) 3) C190S-MRP1 2.9 ( 0.4 (0.8) (n ) 4) 2.7 ( 0.4 (1.4) (n ) 3) C208A-MRP1 3.0 ( 0.6 (0.8) (n ) 3) 2.4 ( 0.6 (1.2) (n ) 3) C208S-MRP1 4.2 ( 0.8 (1.2) (n ) 3) 3.8 ( 0.7 (1.9) (n ) 3) C265A-MRP1 2.5 ( 0.2 (0.7) (n ) 4) 2.3 ( 0.6 (0.9) (n ) 3) C265S-MRP1 10.2 ( 0.4 (2.8)b (n ) 5) 4.0 ( 1.7 (2) (n ) 3) a The resistance of stably transfected HeLa cells was determined using a tetrazolium-based cytotoxicity assay. The relative resistance factors were obtained by dividing the IC50 values for wild-type or Cys mutant MRP1 transfected cells by the IC50 values for empty vector control transfected cells and were normalized for differences in MRP1 expression levels. Login to comment

PMID: 22511347 [PubMed] Qin L et al: "Effect of multiple cysteine substitutions on the functionality of human multidrug resistance protein 1 expressed in human embryonic kidney 293 cells: identification of residues essential for function."

No. Sentence Comment

64 Name Inserted (ϩ) or Deleted (-) Restriction Enzymes Primer Sequence (5Ј-3Јa 1 C208A N.A. CCCTAATCCCGCCCCAGAGTCCAG 2 C265A -BsmI GGAAGAAGGAAGCCGCCAAGACTAG 3 C375A -PstI GTCACTGCCGCCTTGCAGACCCTCG 4 C388A N.A. CTTCCACATCGCCTTCGTCAGTGG 5 C555A ϩSpoI/NruI CACCTGGGTCGCGACGCCCTTTCTG 6 C563A ϩBalI/MscI GGTGGCCTTGGCCACATTTGCCGTC 7 C682A ϩNarI CCAGGTGGGCGCCGGAAAGTCGTC 8 C730A ϩEspI, SacI ATCCTTTTTGGAGCTCAGCTGGAGG 9 C744A -StuI GATACAGGCCGCTGCCCTCCTCC 10 C984A ϩBalI/MscI TTCCTTTTCATGGCCAACCATGTGTCC 11 C1047A ϩSacI/SstI TTGGCTTCCCGAGCTCTGCACGTGG 12 C1105A ϩPvuI CATTGGTGCCGCGATCGTTATCCTG 13 C1205A N.A. GCGGCTGGAGGCTGTGGGCAACTG 14 C1209A ϩPvuI GTGTGGGCAACGCGATCGTTCTGTTTG 15 C1299A ϩMluI TTCCGGAACTACGCGTTGCGCTACCGAG 16 C1423A ϩPstI CTAGACCATGAAGCTGCAGAAGGC 17 C1439A ϩPflMI CCAGCTTGTGGCCCTAGCCCGGG 18 C1479A N.A. GTTCGAGGACGCCACCGTCCTCAC 19 Rec L N.A. GAAACCATCCACGACCCTAATCCCGCCCCAGAG 20 Rec R N.A. GGATTAGGGTCGTGGATGGTTTCCGAGAACAG 21 KbnL N.A. CATGGTACCATGGCGCTCCGGGGCTTCTGCAGC 22 KbnR N.A. GGCAGGATCCTTGGAGGAGTACACAACCTTC N.A., not applicable. Login to comment
191 The activity of ⌬MRP1204-282cysless containing the C208A and C265A mutations was most dramatically affected and was decreased by ϳ70% (Fig. 5B, top panel). Login to comment
192 Subsequently, we found that LTC4 transport activity of the C208A and C265A double mutant could be substantially recovered by growth of the HEK transfectants at 28°C (Fig. 5B, bottom panel). Login to comment
190 The activity of èc;MRP1204-282cysless containing the C208A and C265A mutations was most dramatically affected and was decreased by b03;70% (Fig. 5B, top panel). Login to comment