ABCMdb

ABCB1 p.Phe1086Cys

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Publications

PMID: 18708637 [PubMed] Loo TW et al: "Processing mutations disrupt interactions between the nucleotide binding and transmembrane domains of P-glycoprotein and the cystic fibrosis transmembrane conductance regulator (CFTR)."

No. Sentence Comment

107 The locations of positions equivalent to cysteines S473C and R905C in the other half of P-gp (A266C/F1086C) are indicated. Login to comment
151 Because no cross-linking studies have identified cysteines that could be cross-linked at the TMD1-NBD2 interface, we constructed a series of double cysteine mutants between a cysteine in TMD1 (A266C or F267C) and another in NBD2 (R1085C, F1086C, Y1087C or D1088C) that would be equivalent to L443C(NBD1) and S909C(TMD2), respectively, in each half of P-gp. Login to comment
164 A, membranes prepared from cells expressing mutant A266C/F1086C were treated with (ϩ) or without (-) 1 mM copper phenanthroline (CuP) for 15 min at 0 °C. Membranes were also treated with drug substrates or ATP plus vanadate as described in the legend to Fig. 4. Login to comment
170 The verapamil-stimulated ATPase activities of P-gp mutants containing only the Cys-266 or Cys-1086 mutation showed that the reduced activity was due to the presence of the F1086C change (data not shown). Login to comment
213 NBD-TMD contacts appear to be critical for function because cross-linking of mutant L443C(NBD1)/S909C(TMD2) (Fig. 3) or introduction of the F1086C mutation at the TMD1/NBD2 interface inhibited activity. Login to comment

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."

No. Sentence Comment

164 Accordingly, we introduced the W232R mutation into mutant A266C/F1086C/L1260A. Login to comment
166 Figure 3C shows that the W232R mutation promoted maturation of the mutant A266C/F1086C/L1260A (lane 3) and that only the mature protein was cross-linked (lane 4). Login to comment
188 Membranes were also prepared from cells expressing the L1260A processing mutant ( W232R containing cysteines in TMD1 and NBD2 (A266C/F1086C) (C). Login to comment

PMID: 23634976 [PubMed] Loo TW et al: "A salt bridge in intracellular loop 2 is essential for folding of human p-glycoprotein."

No. Sentence Comment

13 ICL2 appears to play a key role in coupling NBD1-TMD2 interactions because cysteines introduced into ICL2 (A266C) and NBD2 (F1086C) could be cross-linked and the F1086C change abolished activity.13 While both structures predict that residues 261-267 form an interhelical loop (IH2) (forms the ball portion of the ball-and-socket ICL2-NBD connection), adjacent amino acids were predicted to adopt loop or b1;-helical structures in the mouse or C. elegans structures, respectively. Login to comment

PMID: 23733192 [PubMed] Loo TW et al: "Human P-glycoprotein contains a greasy ball-and-socket joint at the second transmission interface."

No. Sentence Comment

56 IH2 appears to be particularly important because we showed that a cysteine introduced into IH2 (A266C) could be directly cross-linked to a cysteine in NBD2 (F1086C) but the mutant was inactive (21). Login to comment
62 Because mutant A266C/F1086C was inactive, we characterized the Ala-266/ Phe-1086 interface to determine its role in the transport cycle. Login to comment
109 In a previous study, we provided biochemical evidence that Ala-266 was indeed close to Phe-1086 in NBD2 because mutant A266C/F1086C showed robust cross-linking even when treated with oxidant at 0 &#b0;C to slow molecular motion (21). Login to comment
110 The Ala-266/Phe-1086 contact point appeared to be critical for function because mutant A266C/F1086C was inactive (21). Login to comment
117 IH2 Mediates TMD/NBD Coupling in the P-gp Drug Pump 20328 JOURNAL OF BIOLOGICAL CHEMISTRY VOLUME 288ߦNUMBER 28ߦJULY 12, 2013 To determine whether one or both cysteine mutations in mutant A266C/F1086C inhibited activity, mutants were constructed in a Cys-less background that contained only Cys-266 or Cys-1086. Login to comment
119 It was observed that only the F1086C mutation caused a drastic reduction in verapamil-stimulated ATPase activity (Fig. 3C). Login to comment
120 An explanation for the ability of the F1086C mutation to inhibit P-gp activity was that the mutation affected folding of P-gp. Login to comment
121 To test whether the F1086C mutation inhibited folding, the F1086C mutant and the Y1087C mutant were expressed in the absence of drug substrates. Login to comment
123 It was found that both the F1086C and the Y1087C mutations inhibited maturation of Cys-less P-gp (Fig. 3D). Login to comment
125 In addition, it was possible that replacement of Phe-1086 with a cysteine caused a defect in the structure of P-gp that would be different if it had been replaced with a smaller amino acid. Login to comment
138 A and B, structures of P-gp in the open (A) (17) or closed (B) (24) conformations are shown. C, histidine-tagged Cys-less P-gp or mutants A266C/F1086C, A266C, and F1086C (in Cys-less background) as well as wild-type P-gp and mutant F1086A ( in wild-type background) were isolated, and ATPase activities were measured in the presence of verapamil. Login to comment

PMID: 25987565 [PubMed] Loo TW et al: "The Transmission Interfaces Contribute Asymmetrically to the Assembly and Activity of Human P-glycoprotein."

No. Sentence Comment

301 For example, the F1086C mutation inhibits maturation of Cys-less P-gp but not wild-type P-gp (23). Login to comment

PMID: 26507655 [PubMed] Loo TW et al: "Mapping the Binding Site of the Inhibitor Tariquidar That Stabilizes the First Transmembrane Domain of P-glycoprotein."

No. Sentence Comment

65 Membranes were prepared and samples were incubated at 0 &#b0;C (A80C/R741C, A259C/W803C, I299C/F770C, and A266C/ F1086C) or 20 &#b0;C (L175C/N820C, C431/L1176C, and L521C/C1074) for 10 min in the presence or absence of 0.5 Mapping the P-glycoprotein Tariquidar-binding Site 29390 JOURNAL OF BIOLOGICAL CHEMISTRY VOLUME 290ߦNUMBER 49ߦDECEMBER 4, 2015 at SEMMELWEIS UNIV OF MEDICINE on December 11, mM copper phenanthroline (oxidant to promote disulfide bond formation) in the presence or absence of 0.25 òe;M (for TM segment cysteine mutants) or 1 òe;M tariquidar (for the ICL and NBD cysteine mutants). Login to comment