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Fluorescein derivatives potentiate CFTR by altering the ATP affinity and efficacy of channel gating

Fluorescein derivatives potentiate CFTR by altering the ATP ...

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2008北京国际生理科学学术大会

作者： MORAN Oscar SHEPPARD David N Istituto di Biofisica CNRVia DeMarini 616149 GenoaItaly Department of Physiology and Pharmacology University of BristolBristolBS8 1TDUK

The fluorescein derivative phloxine B is a potent modulator of the cystic fibrosis transmembrane conductance regulator(CFTR)Cl *** micromolar concentrations of phloxine B potentiate ATP-dependent channel gating,whereas higher concentrations occlude the channel *** determine whether fluorescein derivatives (FDs)affect the ATP affinity of CFTR and the efficacy of channel gating,we investigated the effects of FDs on the gating behaviour of CFTR using excised inside-out membrane patches from C127 cells expressing wild-type human *** employed a simple linear three-state scheme(C(?)C(?)O)to describe CFTR channel *** this scheme,C represents the long duration closed state separating channel openings,C the brief closures that interrupt channel openings,and O the open *** between the three states are described by the forward rate constantsβ,β and the backward rate constantsα andα.Given that CFTR is an agonist-activated channel gated by intracellular ATP and this scheme is reminiscent of the del Castillo-Katz mechanism,we can calculate the ATP affinity(α/β)and gating efficacy of CFTR(β/α).The FDs phloxine B(1μmol/L)and eosin Y(1μmol/L) augment channel gating by prolonging mean burst duration(MBD)without changing the interburst interval(IBI), whereas dichlorofluorescein(DCF;20μmol/L)decreases the IBI without changing MBD(n=4-15).Interestingly, phloxine B(1μmol/L)and DCF(20μmol/L)decreasedα/β(enhanced affinity),whereas only phloxine B(1μmol/L) elevatedβ/α(augmented gating efficacy).To understand better how FDs affect the relationship between the ATP affinity and gating efficacy,we examined the ATP-dependence of channel gating(n>5).Increasing ATP concentrations enhanced markedly both the ATP affinity and gating efficacy of ***,phloxine B(1μmol/L) augmented both the ATP affinity and gating efficacy of CFTR,particularly at low ATP *** interpret our results to suggest that the ATP affinity and gating efficacy of CFTR are t

关键词： CFTR channel gating ATP affinity gating efficacy

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