ABCMdb

PMID: 16624245

Ghosh P, Moitra K, Maki N, Dey S
Allosteric modulation of the human P-glycoprotein involves conformational changes mimicking catalytic transition intermediates.
Arch Biochem Biophys. 2006 Jun 1;450(1):100-12. Epub 2006 Mar 15., [PubMed]

Sentences

No. Mutations Sentence Comment

3 ABCB1 p.Phe983Ala The protection by Xupentixol is abolished in the Pgp F983A mutant that is impaired in modulation by Xupentixols, indicating involvement of the allosteric site in generating the conformational change. Login to comment 46 ABCB1 p.Phe983Ala Baculovirus mediated expression of human Pgp A recombinant baculovirus harboring either the wild-type or the F983A mutant human MDR1 cDNA, with 6 £His-tag at the C-terminal end, BV-MDR1-(H6) [34] was used to infect High Five insect cells grown in serum free Excell 400 medium as described [35]. Login to comment 105 ABCB1 p.Phe983Ala SigniWcantly, no detectable protection from trypsin digestion was observed for the Pgp F983A mutant (Fig. 2C) that is impaired in modulation by Xupentixol [21]. Login to comment 107 ABCB1 p.Phe983Ala SigniWcantly, no detectable protection from trypsin digestion was observed for the Pgp F983A mutant (Fig. 2C) that is impaired in modulation by Xupentixol [21]. Login to comment 112 ABCB1 p.Phe983Ala Protection to limited trypsin digestion is induced by both isomers of Xupentixol, which is aVected in Pgp F983A mutant. Login to comment 114 ABCB1 p.Phe983Ala Protection to limited trypsin digestion is induced by both isomers of Xupentixol, which is aVected in Pgp F983A mutant. Login to comment 116 ABCB1 p.Phe983Ala (C) Similar experiments were carried out with the Pgp F983A mutant in the presence of varying concentrations (0-25 M) of cis-(Z)-Xupentixol and resolved as stated above. Login to comment 117 ABCB1 p.Phe983Ala 198 49.8 115 93 +-Trypsin 0 1 2.5 5 10cis-(Z)-Flupentixol (µM) 250 Wild Type 198 49.8 115 93 +-Trypsin 0 1 2.5 5 10trans-(E)-Flupentixol (µM) 250 Wild Type 198 49.8 115 93 +-Trypsin 0 1 2.5 5 10cis-(Z)-Flupentixol (µM) 250 F983A A B C strate site, both isomers of Xupentixol induced similar conformational changes in Pgp. Login to comment 118 ABCB1 p.Phe983Ala (C) Similar experiments were carried out with the Pgp F983A mutant in the presence of varying concentrations (0-25 M) of cis-(Z)-Xupentixol and resolved as stated above. Login to comment 119 ABCB1 p.Phe983Ala 198 49.8 115 93 + - Trypsin 0 1 2.5 5 10 cis-(Z)-Flupentixol (&#b5;M) 25 0 Wild Type 198 49.8 115 93 + - Trypsin 0 1 2.5 5 10 trans-(E)-Flupentixol (&#b5;M) 25 0 Wild Type 198 49.8 115 93 + - Trypsin 0 1 2.5 5 10 cis-(Z)-Flupentixol (&#b5;M) 25 0 F983A A B C strate site, both isomers of Xupentixol induced similar conformational changes in Pgp. Login to comment 188 ABCB1 p.Phe983Ala Since the Pgp F983A mutant that is impaired in modulation by Xupentixol [21,22] displays no such protection (Fig. 2C) the functional signiWcance of the conformational change in the mechanism of modulation is underscored. Login to comment 190 ABCB1 p.Phe983Ala Since the Pgp F983A mutant that is impaired in modulation by Xupentixol [21,22] displays no such protection (Fig. 2C) the functional signiWcance of the conformational change in the mechanism of modulation is underscored. Login to comment 223 ABCB1 p.Phe983Ala Consistent with that, in the Pgp F983A mutant, the loss of cis-(Z)-Xupentixol-induced conformational change coincides with impaired modulation of [125 I]IAAP binding, whereas inhibition of [125 I]IAAP binding by cyclosporin A remained unaltered in the mutant [21]. Login to comment 225 ABCB1 p.Phe983Ala Consistent with that, in the Pgp F983A mutant, the loss of cis-(Z)-Xupentixol-induced conformational change coincides with impaired modulation of [125 I]IAAP binding, whereas inhibition of [125 I]IAAP binding by cyclosporin A remained unaltered in the mutant [21]. Login to comment