Similar(8)
No inward current was detectable when Na+ in the bath solution was substituted with choline, indicating that the channel mutant was Ca2+ impermeable (Erev = 16.7 ± 1.1 mV, n = 14; Figures 1I 1K).
After it was established that most mutants were functional, the effect of the negatively charged and membrane impermeable MTSES on wild-type OATP1B1 and its different cysteine mutants was examined.
The aim of the present study was to clarify the location of the diltiazem binding site making use of its membrane-impermeable quaternary derivative d- cis-diltiazem (qDil) and mutant α1 subunits.
Cell surface expression of the D50A and T118A mutant receptors was confirmed using a membrane-impermeable biotinylating agent as described elsewhere [1] (data not shown).
To test whether the observed effects were due to the negative charge introduced by MTSES or a steric effect of MTSES, we treated cells expressing wild-type OATP1B1 and certain mutants with the positively charged and membrane impermeable MTSET.
Cell surface expression of the mutant D50A and T118A ANTXreceptorsors was confirmed using a membrane-impermeable biotinylating reagent (EZ-Link SuLFo-NHS-Biotin; Pierce) and pelleting biotinylated proteins with immobilized streptavidin gel (Immunopure; Pierce) essentially as described elsewhere [1].
Mutant Games?
An impermeable police hierarchy is another problem.
Related(1)
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