ABCC1 p.Asp792Ala
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PMID: 11741902
[PubMed]
Hou YX et al: "ATP binding to the first nucleotide-binding domain of multidrug resistance protein MRP1 increases binding and hydrolysis of ATP and trapping of ADP at the second domain."
No.
Sentence
Comment
50
Stable cell lines expressing wild-type and mutant MRP1s, K684L, D792A, K1333L, and D1454L/E1455L were established previously (2, 31).
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ABCC1 p.Asp792Ala 11741902:50:64
status: NEW117 Essentially the stimulation was much reduced in the NBD1 mutants, K684L and D792A (Fig. 4, C and D), and in the NBD2 mutants, K1333L and D1454L/E1455L (Fig. 4, E and F), and the stimulation effects were shifted to higher ATP concentrations (Fig. 4, C-F).
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ABCC1 p.Asp792Ala 11741902:117:76
status: NEW173 Lane 1, 10 g of wild-type MRP1; lane 2, 15 g of K684L; lane 3, 20 g of D792A; lane 4, 10 g of K1333L; lane 5, 10 g of D1454L/E1455L.
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ABCC1 p.Asp792Ala 11741902:173:95
status: NEW175 The results for K684L and D792A are the average of three independent experiments and for K1333L and D1454L/E1455L are the average of two independent experiments. C, influence of ATP on the [␣-32 P]8-N3ADP labeling of K684L.
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ABCC1 p.Asp792Ala 11741902:175:26
status: NEW176 15 g of K684L was labeled in the presence of varying amounts of ATP indicated above each lane. D, influence of ATP on the [␣-32 P]8-N3ADP labeling of D792A.
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ABCC1 p.Asp792Ala 11741902:176:165
status: NEW177 20 g of D792A was labeled in the presence of varying amount of ATP indicated above each lane.
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ABCC1 p.Asp792Ala 11741902:177:16
status: NEW
PMID: 12458196
[PubMed]
Hou YX et al: "ATP binding, not hydrolysis, at the first nucleotide-binding domain of multidrug resistance-associated protein MRP1 enhances ADP.Vi trapping at the second domain."
No.
Sentence
Comment
226
Indeed, mutations of K684L and D792A greatly diminish the ATP enhancing effect on ADP trapping (34).
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ABCC1 p.Asp792Ala 12458196:226:31
status: NEW
PMID: 16442101
[PubMed]
Frelet A et al: "Insight in eukaryotic ABC transporter function by mutation analysis."
No.
Sentence
Comment
161
The variant D792L was unable to mature conformationally and D792A led to an accumulation of equal amounts of mature and immature proteins but still resulted in defective nucleotide interaction and organic anion transport, indicating that nucleotide hydrolysis at NBD1 was essential for MRP1 function [101].
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ABCC1 p.Asp792Ala 16442101:161:60
status: NEW
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
21
However, substitution of the acidic amino acid D792 in Walker B motif with a hydrophobic residue, such as D792L- or D792A-mutated MRP1 [20,23], caused misfolding of the protein and prevented further analysis of the mutated protein.
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ABCC1 p.Asp792Ala 18088596:21:116
status: NEW24 However, this mechanism of protein folding may not be applied to the misfolding caused by substitution of the acidic amino acid with a hydrophobic residue, such as D792L- or D792A-mutated MRP1 [20,23].
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ABCC1 p.Asp792Ala 18088596:24:174
status: NEW27 We suspected that D792A or D792L mutation abolished the hydrogen-bond formation between D792 and S685 and resulted in misfolding of the mutated MRP1 protein.
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ABCC1 p.Asp792Ala 18088596:27:18
status: NEW94 We suspected that D792A or D792L mutation abolished the hydrogen-bond formation between D792 and S685 and resulted in misfolding of the mutated MRP1 protein.
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ABCC1 p.Asp792Ala 18088596:94:18
status: NEW
PMID: 11469806
[PubMed]
Cui L et al: "Mutations of the Walker B motif in the first nucleotide binding domain of multidrug resistance protein MRP1 prevent conformational maturation."
No.
Sentence
Comment
8
A different substitution of the same residue (D792A) had a less severe effect enabling accumulation of approximately equal amounts of mature and immature MRP1 proteins in the membrane vesicles but still resulted in defective nucleotide interaction and organic anion transport, indicating that nucleotide hydrolysis at NBD1 is essential to MRP1 function.
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ABCC1 p.Asp792Ala 11469806:8:46
status: NEW18 A different substitution of the important residue (D792A) only partially impaired maturation yet still 1 To whom correspondence and reprint requests should be addressed.
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ABCC1 p.Asp792Ala 11469806:18:51
status: NEW36 The aspartic acid residues at positions of 792 and 793 were mutated either to alanine (Fig. 1B, D792A) or leucine residues (Fig. 1B, D792L and D793L) using the QuikChange Site Directed Mutagenesis kit.
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ABCC1 p.Asp792Ala 11469806:36:96
status: NEW43 The cell lines expressing D792A, D792L, and D793L were generated using the same procedures (9).
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ABCC1 p.Asp792Ala 11469806:43:26
status: NEW90 D793L is indistinguishable from wild-type, whereas D792L is similar to the double mutant indicating that the substitution at this position is primarily responsible for the misprocessing.
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ABCC1 p.Asp792Ala 11469806:90:52
status: NEW91 However, when this residue was replaced by alanine (D792A) instead of leucine a detectable mature band was also present.
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ABCC1 p.Asp792Ala 11469806:91:52
status: NEW93 These were more apparent in a longer exposure of the same blot (Fig. 2B), especially in D792L/D793L, where, in addition to the major 170-kDa species, bands of approximately 160, 130, 100, and 30 kDa can be seen.
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ABCC1 p.Asp792Ala 11469806:93:61
status: NEW94 Of these four only the 130- and 30-kDa bands are seen in the D792A and D792L lanes.
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ABCC1 p.Asp792Ala 11469806:94:61
status: NEW99 It is also apparent that the mature form of MRP1 is enriched in the membrane fraction compared with the whole cell lysate; this is especially obvious with D792A, where there appears to be about equal amounts of the immature and mature bands in the membrane fraction (Fig. 2C).
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ABCC1 p.Asp792Ala 11469806:99:155
status: NEW124 The following amounts of protein were loaded in each lane: 0.5 g of wild-type MRP1; 8 g of D792A; 18 g of D792L; 0.5 g of D793L; 20 g of D792L/D793L.
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ABCC1 p.Asp792Ala 11469806:124:107
status: NEW132 The following amounts were loaded in each lane: 0.5 g of wild-type MRP1; 2 g of D792A; 4 g of D792L; 0.35 g of D793L; 8 g of D792L/D793L.
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ABCC1 p.Asp792Ala 11469806:132:64
status: NEWX
ABCC1 p.Asp792Ala 11469806:132:96
status: NEW133 The ratio of mature to immature protein in membrane vesicles of D792A and D792L are much higher than in whole cell lysates (Fig. 2A).
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ABCC1 p.Asp792Ala 11469806:133:64
status: NEW136 Lanes 1 and 2, 1 g of D793L cell lysates in each lane; Lanes 3 and 4, 4 g of D792A cell lysates in each lane; Lanes 5 and 6, 10 g of D792L/D793L cell lysates in each lane.
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ABCC1 p.Asp792Ala 11469806:136:67
status: NEWX
ABCC1 p.Asp792Ala 11469806:136:93
status: NEW137 Both the 170-kDa core-glycosylated MRP1 protein from either D793L, D792A, or D792L/D793L and 160-kDa degradation product from D792L/D793L were decreased in size by treatment with endoglycosidase H. pressing either the D792L or D792L/D793L mutants with either lactacystin or ALLN resulted in the total disappearance of the immature forms from nonionic detergent soluble fractions and appearance in insoluble pellets.
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ABCC1 p.Asp792Ala 11469806:137:67
status: NEW155 This comparison is most obvious with the D792A mutant where the membranes subjected to trypsin digestion contain approximately equal amounts of immature and mature species (Fig. 5C).
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ABCC1 p.Asp792Ala 11469806:155:41
status: NEW168 It was impossible to tell if the dysfunction of the D792L mutation was secondary to its inability to mature conformationally.
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ABCC1 p.Asp792Ala 11469806:168:41
status: NEW169 However, since approximately half of the D792A protein in the membrane was mature it was possible to assay its functional capability.
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ABCC1 p.Asp792Ala 11469806:169:41
status: NEW171 Figure 6 shows labeling of the wild-type and the innocuous D793L variant which matures normally.
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ABCC1 p.Asp792Ala 11469806:171:100
status: NEW172 However, there was greatly reduced labeling of the variants in which D792 was substituted including D792A where there is considerable mature protein.
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ABCC1 p.Asp792Ala 11469806:172:100
status: NEW174 Since hydrolysis is believed to drive MRP1 transport it would be expected that the mature D792A protein would not be capable of active transport.
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ABCC1 p.Asp792Ala 11469806:174:90
status: NEWX
ABCC1 p.Asp792Ala 11469806:174:132
status: NEW175 The data in Fig. 7 confirm this expectation, i.e., there is not significantly more ATP-dependent LTC4 uptake by vesicles containing D792A protein that does mature than by the other variants that do not mature (Fig. 7, D792L and D792L/D793L), nor by the NBD2 mutants (Fig. 7, K1333L and D1454L/E1455L) that do mature but have difficulties to hydrolyze ATP and to trap the hydrolysis product, ADP (8).
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ABCC1 p.Asp792Ala 11469806:175:132
status: NEW184 However, when a different substitution, D792A, was made considerable maturation occurred although still less than wild-type (Figs. 2B and 2C).
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ABCC1 p.Asp792Ala 11469806:184:40
status: NEW203 (B) K684L, 0.6 g protein in each lane.
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ABCC1 p.Asp792Ala 11469806:203:4
status: NEW204 (C) D792A, 0.73 g protein in each lane.
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ABCC1 p.Asp792Ala 11469806:204:4
status: NEW98 It is also apparent that the mature form of MRP1 is enriched in the membrane fraction compared with the whole cell lysate; this is especially obvious with D792A, where there appears to be about equal amounts of the immature and mature bands in the membrane fraction (Fig. 2C).
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ABCC1 p.Asp792Ala 11469806:98:155
status: NEW123 The following amounts of protein were loaded in each lane: 0.5 òe;g of wild-type MRP1; 8 òe;g of D792A; 18 òe;g of D792L; 0.5 òe;g of D793L; 20 òe;g of D792L/D793L.
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ABCC1 p.Asp792Ala 11469806:123:105
status: NEW131 The following amounts were loaded in each lane: 0.5 òe;g of wild-type MRP1; 2 òe;g of D792A; 4 òe;g of D792L; 0.35 òe;g of D793L; 8 òe;g of D792L/D793L.
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ABCC1 p.Asp792Ala 11469806:131:94
status: NEW135 Lanes 1 and 2, 1 òe;g of D793L cell lysates in each lane; Lanes 3 and 4, 4 òe;g of D792A cell lysates in each lane; Lanes 5 and 6, 10 òe;g of D792L/D793L cell lysates in each lane.
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ABCC1 p.Asp792Ala 11469806:135:91
status: NEW154 This comparison is most obvious with the D792A mutant where the membranes subjected to trypsin digestion contain approximately equal amounts of immature and mature species (Fig. 5C).
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ABCC1 p.Asp792Ala 11469806:154:41
status: NEW173 Since hydrolysis is believed to drive MRP1 transport it would be expected that the mature D792A protein would not be capable of active transport.
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ABCC1 p.Asp792Ala 11469806:173:90
status: NEW183 However, when a different substitution, D792A, was made considerable maturation occurred although still less than wild-type (Figs. 2B and 2C).
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ABCC1 p.Asp792Ala 11469806:183:40
status: NEW