ABCB1 p.Trp232Ala

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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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58 (B) HEK 293 cells were transfected with A52-tagged wild-type P-gp, mutants G251V, G251V/W232R, and G251V/ W232A,orplasmidvector(control).Wholecellextractsweresubjected to immunoblot analysis with monoclonal antibody against A52 or GAPDH.
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ABCB1 p.Trp232Ala 21182301:58:106
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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.Trp232Ala 21182301:84:107
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114 We tested whether a W232A change would also promote maturation of the G251V mutant.
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ABCB1 p.Trp232Ala 21182301:114:20
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118 By contrast, immature protein was the major product in mutants G251V (10 ( 4% mature) and G251V/W232A (13 ( 5% mature).
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ABCB1 p.Trp232Ala 21182301:118:96
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285 Stable BHK cell lines expressing mutants W232A, N296A, E875A, or T945A were generated.
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ABCB1 p.Trp232Ala 21182301:285:41
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289 For example, cells expressing mutants W232A and N296A were more resistant to colchicine (P < 0.05) than cells expressing wild-type P-gp, but their resistance to paclitaxel was reduced (P < 0.05).
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ABCB1 p.Trp232Ala 21182301:289:38
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299 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.Trp232Ala 21182301:299:144
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304 The verapamil-stimulated ATPase activities of mutants W232A and N296A, however, resembled that of wild-type enzyme (S50 of 30 and 42 μM, respectively).
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ABCB1 p.Trp232Ala 21182301:304:54
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307 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.Trp232Ala 21182301:307:45
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308 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.Trp232Ala 21182301:308:19
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310 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.Trp232Ala 21182301:310:117
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336 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.Trp232Ala 21182301:336:37
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