ABCMdb

ABCC1 p.Asp1454Asn

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Publications

PMID: 15755910 [PubMed] Payen L et al: "Functional interactions between nucleotide binding domains and leukotriene C4 binding sites of multidrug resistance protein 1 (ABCC1)."

No. Sentence Comment

67 The forward primers for the D792N and D1454N mutations of the Walker B motifs were 5Ј-CATTTACCTCT- TCAATGATCCCCTC-3Ј and 5Ј-ATCCTTGTGTTGAATGAGGCCA- CG-3Ј, respectively. Login to comment
189 The D792N mutation diminished transport activity by approximately 65%, whereas the D1454N mutation essentially inactivated the protein (Fig. 5B). Login to comment
191 In contrast, the D1454N mutation eliminated photolabeling of only NBD2 and had no effect on photolabeling of NBD1 (Fig. 5C). Login to comment
192 The D792N mutation also strongly decreased ADP trapping by both NBD1 and NBD2, although the D1454N mutation eliminated trapping by NBD2 but only modestly decreased trapping by NBD1 (Fig. 6B). Login to comment
228 The relative expression levels of wt and mutant half-molecules were evaluated by densitometry and are indicated on the figure. B, effect of D792N and D1454N mutations on ATP-dependent LTC4 transport activity. Login to comment
229 Membrane vesicles (2 ␮g) containing wt and the D792N and D1454N mutant MRP1 half-molecules or control beta-gus were assayed for ATP-dependent LTC4 transport activity by incubation in transport buffer containing [3 H]LTC4 (50 nM, 0.13 ␮Ci) at 23°C for 2 min in the presence and absence of ATP (4 mM) as described under Materials and Methods. Results shown are means Ϯ S.D. of triplicate determinations in a single experiment. Login to comment
230 Similar results were obtained in three additional independent experiments. C, effect of D792N and D1454N mutations on photolabeling with 8-azido-[␣-32 P]ATP. Login to comment
242 The effect of the MRP1 Walker B D792N and D1454N mutations on ATP binding and ADP trapping was similar to that of the Walker A lysine mutations; the level of transport activity of the NBD1 aspartic acid-to-asparagine mutation was comparable with that of the Walker A lysine-to-methionine mutation (Gao et al., 2000). Login to comment
258 Densitometry indicated that the level of the D1454N mutant protein in the vesicle preparation used was approximately 2-fold higher than in control vesicles (A). Login to comment
260 B, effect of D792N and D1454N mutations on vanadate-dependent nucleotide trapping. Login to comment
265 C, effect of D792N and D1454N mutations on LTC4 photolabeling in the presence or absence of nucleotide. Login to comment
266 Membrane vesicles (50 ␮g of total protein) containing wt and the D792N and D1454N mutant MRP1 half-molecules were incubated in transport buffer at 23°C for 20 min in the absence or presence of ATP␥S (4 mM) or ATP (1 mM) plus vanadate (1 mM) before the addition of [3 H]LTC4 (200 nM, 0.13 ␮Ci). Login to comment

PMID: 17187755 [PubMed] Yang R et al: "Hydrogen-bond formation of the residue in H-loop of the nucleotide binding domain 2 with the ATP in this site and/or other residues of multidrug resistance protein MRP1 plays a crucial role during ATP-dependent solute transport."

No. Sentence Comment

189 For example, ATP binding to wild-type MRP1 can transport the bound LTC4 from high to low affinity site whereas ATP binding to the K684R- or the D1454N-mutated MRP1 cannot [30]. Login to comment

PMID: 17295059 [PubMed] Chang XB et al: "A molecular understanding of ATP-dependent solute transport by multidrug resistance-associated protein MRP1."

No. Sentence Comment

241 Indeed, several mutations, such as K684E, K1333E, K684R, K1333R, D792N, D1454N, G771A and G1433A, significantly diminished ATP binding and Vi-dependent ADP trapping at NBD2 and lost the ability to shift the substrate binding from a high to low affinity site [61]. Login to comment

PMID: 18088596 [PubMed] Yang R et al: "The hydroxyl group of S685 in Walker A motif and the carboxyl group of D792 in Walker B motif of NBD1 play a crucial role for multidrug resistance protein folding and function."

No. Sentence Comment

254 In order to rule out the possibility that the double mutant D1454L/E1455L might rescue the misfolding caused by D14 54L mutation, we have made single mutants including D1454L, D1454N, S1334A, S1334T, S1334C, S1334H, S1334D and S1334N. Login to comment
261 It is also true for the corresponding mutations in Walker B motif of NBD2 in MRP1, such as D1454L (manuscript in preparation) and D1454N [46], have a significant effect on the ATP-dependent LTC4 transport. Login to comment

PMID: 19949927 [PubMed] Chang XB et al: "Molecular mechanism of ATP-dependent solute transport by multidrug resistance-associated protein 1."

No. Sentence Comment

157 Indeed, several mutations, such as K684E, K1333E, K684R, K1333R, D792N, D1454N, G771A and G1433A, significantly diminished ATP binding and lost the ability to shift the bound substrate from high to low affinity site (99). Login to comment

PMID: 21799180 [PubMed] Henderson MJ et al: "ABCC multidrug transporters in childhood neuroblastoma: clinical and biological effects independent of cytotoxic drug Efflux."

No. Sentence Comment

66 For constitutive expression of ABCC1 in SH-SY5Y cells, transfection with pCMV14-3xFLAG-ABCC1, or the ATP-binding site mutants, pCMV14-3xFLAG-ABCC1-D1454N or ABCC1-DE1454LL, was followed by selection of stable clones with 500 µg/ml G418 (Geneticin; Invitrogen Australia Pty Ltd, Mulgrave, Australia), which were subsequently maintained with 150 µg/ml G418. Login to comment
67 The ABCC1-D1454N mutation was previously shown to abolish ATP-dependent ABCC1 transporter activity while not affecting correct localization of the transporter (19), and the ABCC1-DE1454LL mutant was predicted to have similar properties (Supplementary Figure 1, available online and http://www.uniprot.org/uniprot/O15439; accessed June, 2010). Login to comment
74 Generation of ATP-binding site mutants was carried out accordingtotheQuikChangeLightningSite-DirectedMutagenesis Kit (Stratagene, La Jolla, CA) using forward primer 5'-ATC CTT GTG TTG AAT GAG GCC ACG GCA G-3' for ABCC1-D1454N (single mutant), forward primer 5'-ATC CTT GTG TTG CTT CTG GCC ACG GCA G-3' for ABCC1-DE1454/1455LL (double mutant) and forward primer 5'-AGG TGG GGA TCT TCG GCC GCA CTG G-3' for ABCC3-V1322F. Login to comment
151 ABCC1 transporter activity is necessary for the observed effects on cell motility, because forced expression of catalytically inactive ABCC1 (ABCC1-D1454N clone B4 or ABCC1-DE1454LL clone B7) had no impact on cell motility (mean % of wound open ± 95% CI: ABCC1 B4, 49.6 ± 4.5 vs empty vector, 59.9 ± 9.6, P = .08; ABCC1 B7, 55.6 ± 9.0 vs empty vector, 59.9 ± 9.6, P = .54, one-way ANOVA and two-sided t test, three independent Figure 2.Impact of ABCC1 and ABCC4 suppression in BE(2)-C human neuroblastoma cells. Login to comment
163 This effect was completely abrogated in cells overexpressing the mutant ABCC1 carrying a single amino acid change in the ABC domain (D1454N) or the catalytically inactive double mutant of ABCC1 (DE1454LL) (mean number of colonies ± 95% CI: ABCC1 B4, 74.8 ± 1.6 vs empty vector, 73.9 ± 4.3, P = .96; ABCC1 B7, 73.5 ± 5.9 vs empty vector, 73.9 ± 4.3, P = .88; one-way ANOVA and two-sided t test, four independent experiments; Figure 3, D). Login to comment
182 A) Western blot analysis of ABCC1 protein expression following stable transduction of SH-SY5Y cells with either empty vector, wild-type (wt) ABCC1 (clones D5, H7), ABCC1 D1454N single mutant (clone B4), or ABCC1 DD1454LL double mutant (clone B7) constructs. Login to comment
68 The ABCC1-D1454N mutation was previously shown to abolish ATP-dependent ABCC1 transporter activity while not affecting correct localization of the transporter (19), and the ABCC1-DE1454LL mutant was predicted to have similar properties (Supplementary Figure 1, available online and http://www.uniprot.org/uniprot/O15439; accessed June, 2010). Login to comment
75 Generation of ATP-binding site mutants was carried out accordingtotheQuikChangeLightningSite-DirectedMutagenesis Kit (Stratagene, La Jolla, CA) using forward primer 5'-ATC CTT GTG TTG AAT GAG GCC ACG GCA G-3' for ABCC1-D1454N (single mutant), forward primer 5'-ATC CTT GTG TTG CTT CTG GCC ACG GCA G-3' for ABCC1-DE1454/1455LL (double mutant) and forward primer 5'-AGG TGG GGA TCT TCG GCC GCA CTG G-3' for ABCC3-V1322F. Login to comment
152 ABCC1 transporter activity is necessary for the observed effects on cell motility, because forced expression of catalytically inactive ABCC1 (ABCC1-D1454N clone B4 or ABCC1-DE1454LL clone B7) had no impact on cell motility (mean % of wound open ± 95% CI: ABCC1 B4, 49.6 ± 4.5 vs empty vector, 59.9 ± 9.6, P = .08; ABCC1 B7, 55.6 ± 9.0 vs empty vector, 59.9 ± 9.6, P = .54, one-way ANOVA and two-sided t test, three independent Figure 2.Impact of ABCC1 and ABCC4 suppression in BE(2)-C human neuroblastoma cells. Login to comment
164 This effect was completely abrogated in cells overexpressing the mutant ABCC1 carrying a single amino acid change in the ABC domain (D1454N) or the catalytically inactive double mutant of ABCC1 (DE1454LL) (mean number of colonies ± 95% CI: ABCC1 B4, 74.8 ± 1.6 vs empty vector, 73.9 ± 4.3, P = .96; ABCC1 B7, 73.5 ± 5.9 vs empty vector, 73.9 ± 4.3, P = .88; one-way ANOVA and two-sided t test, four independent experiments; Figure 3, D). Login to comment
183 A) Western blot analysis of ABCC1 protein expression following stable transduction of SH-SY5Y cells with either empty vector, wild-type (wt) ABCC1 (clones D5, H7), ABCC1 D1454N single mutant (clone B4), or ABCC1 DD1454LL double mutant (clone B7) constructs. Login to comment