ABCB1 p.Thr199Cys
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PMID: 17848563
[PubMed]
Loo TW et al: "Suppressor mutations in the transmembrane segments of P-glycoprotein promote maturation of processing mutants and disrupt a subset of drug-binding sites."
No.
Sentence
Comment
147
Fig. 2A shows that all of the mutants except T199C had activities similar to that of Cys-less P-gp (less than 1.5-fold increase).
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ABCB1 p.Thr199Cys 17848563:147:45
status: NEW148 Mutant T199C showed a 6.7-fold increase in activity after treatment with MTS-rhodamine.
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ABCB1 p.Thr199Cys 17848563:148:7
status: NEW154 The mutants Q195C and T199C, however, exhibited different properties than Cys-less P-gp after treatment with MTS-rhodamine.
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ABCB1 p.Thr199Cys 17848563:154:22
status: NEW157 By contrast, covalent modification of mutant T199C with MTS-rhodamine fully activated its ATPase activity such that rhodamine B had no further effect on its activity.
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ABCB1 p.Thr199Cys 17848563:157:45
status: NEW158 If MTS-rhodamine activates P-gp ATPase activity because it occupies the rhodamine-binding site when it is attached to Cys199 , then labeling of mutant T199C should be protectable with rhodamine B.
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ABCB1 p.Thr199Cys 17848563:158:151
status: NEW160 Histidine-tagged mutant T199C was expressed in HEK 293 cells, solubilized with detergent, and reacted with various concentrations of MTS-rhodamine.
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ABCB1 p.Thr199Cys 17848563:160:24
status: NEW162 To test whether rhodamine B protected mutant T199C from labeling, HEK 293 cells expressing the histidine-tagged mutant were solubilized with detergent and treated with 1 mM MTS-rhodamine in the presence or absence of 5 mM rhodamine B for 10 min at 20 °C.
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ABCB1 p.Thr199Cys 17848563:162:45
status: NEW163 The P-gps were isolated by nickel-chelate chromatography and mixed with lipids, and ATPase activities were determined. Fig. 3B shows that ATP hydrolysis was reduced by 78% when mutant T199C was reacted with MTS-rhodamine in the presence of 5 mM rhodamine B.
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ABCB1 p.Thr199Cys 17848563:163:184
status: NEW168 Fold-stimulation 1 2 3 4 5 6 Cys-less I190C G191C M192C F193C F194C Q195C S196C M197C A198C T199C F200C F201C T202C G203C F204C I205C V206C G207C F208C T209C A 7 Fold-stimulation 1 2 3 4 5 6 Cys-less B _ + MTS-rhod Rhod B+ _ + Q195C + + _ + T199C + + _ + _ _+ + + 7 FIGURE 2.
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ABCB1 p.Thr199Cys 17848563:168:92
status: NEWX
ABCB1 p.Thr199Cys 17848563:168:241
status: NEW173 B, Cys-less, Q195C and T199C P-gp mutants were treated with (ϩ) or without (-) 2 mM MTS-rhodamine and histidine-tagged P-gp isolated by nickel-chelate chromatography. Equivalent amounts of P-gp were mixed with lipid, and ATPase activity was determined in the presence (ϩ) or absence (-) of 2 mM rhodamine B (Rhod B).
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ABCB1 p.Thr199Cys 17848563:173:23
status: NEW177 Labeling of mutant T199C with increasing concentrations of MTS-rhodamine and protection by rhodamine B.
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ABCB1 p.Thr199Cys 17848563:177:19
status: NEW178 A, HEK 293 cells expressing histidine-tagged mutant T199C were solubilized with n-dodecyl-beta-D-maltoside, and insoluble material was removed by centrifugation.
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ABCB1 p.Thr199Cys 17848563:178:52
status: NEW269 The labeling of Cys199 also appeared to be specific for MTS-rhodamine because the ATPase activity of mutant T199C was not activated by MTS-verapamil.
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ABCB1 p.Thr199Cys 17848563:269:108
status: NEW
PMID: 22700974
[PubMed]
Loo TW et al: "The ATPase activity of the P-glycoprotein drug pump is highly activated when the N-terminal and central regions of the nucleotide-binding domains are linked closely together."
No.
Sentence
Comment
255
Covalent labeling of T199C (TM3) with a thiol-reactive derivative of rhodamine increased basal activity 7-fold (58).
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ABCB1 p.Thr199Cys 22700974:255:21
status: NEW248 Covalent labeling of T199C (TM3) with a thiol-reactive derivative of rhodamine increased basal activity 7-fold (58).
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ABCB1 p.Thr199Cys 22700974:248:21
status: NEW