ABCB1 p.Glu875Ala
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PMID: 21182301
[PubMed]
Loo TW et al: "The W232R suppressor mutation promotes maturation of a truncation mutant lacking both nucleotide-binding domains and restores interdomain assembly and activity of P-glycoprotein processing mutants."
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Sentence
Comment
84
Baby hamster kidney (BHK) cells expressing A52-tagged wild-type P-gp or mutants G251V, G251V/W232R, W232R, W232A, N296A, E875A, or T945A were generated as described previously (25).
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ABCB1 p.Glu875Ala 21182301:84:121
status: NEW238 A G251V/T945R/E875A mutant was constructed to test if removal of the negative charge would affect maturation.
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ABCB1 p.Glu875Ala 21182301:238:14
status: NEW239 It was found that T945R no longer promoted maturation when the E875A mutation was present (Figure 7C, lane 4).
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ABCB1 p.Glu875Ala 21182301:239:63
status: NEW285 Stable BHK cell lines expressing mutants W232A, N296A, E875A, or T945A were generated.
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ABCB1 p.Glu875Ala 21182301:285:55
status: NEW299 Each value is average of triplicate assays ( SD. (B) Whole cell extracts of BHK cells expressing vector (control), A52-tagged wild-type, mutant W232A, N296A, E875A, or T945A P-gp were subjected to immunoblot analysis with monoclonal antibody against A52 or GAPDH.
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ABCB1 p.Glu875Ala 21182301:299:158
status: NEW303 Higher concentrations of verapamil were required for half-maximal activation (S50) of ATPase activity for mutants T945A (370 μM) and E875A (>1 mM) compared to wild-type P-gp (48 μM).
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ABCB1 p.Glu875Ala 21182301:303:139
status: NEW307 By contrast, the maximum activity of mutants W232A, E875A, and T945A were reduced relative to wild-type P-gp.
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ABCB1 p.Glu875Ala 21182301:307:52
status: NEW308 The S50 of mutants W232A (45 μM) and T945A (200 μM) differed from wild-type P-gp (102 μM) while those of mutants N296A (78 μM) and E875A (86 μM) were similar.
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ABCB1 p.Glu875Ala 21182301:308:155
status: NEW310 In summary, the E875A and T945A mutations altered both verapamiland rhodamine B-stimulated ATPase activity while the W232A and N296A mutations only affected rhodamine B-stimulated ATPase activity.
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ABCB1 p.Glu875Ala 21182301:310:16
status: NEW336 Histidine-tagged wild-type or mutant W232A, N296A, E875A, or T945A P-gp was expressed in HEK 293 cells and isolated by nickel-chelate chromatography.
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ABCB1 p.Glu875Ala 21182301:336:51
status: NEW