Thiocyanate as a probe of the cystic fibrosis transmembrane conductance regulator chloride channel pore

Abstract

Immediately following exposure to thiocyanate (SCN–)-containing solutions, the cystic fibrosis conductance regulator Cl– channel exhibits high unitary SCN– conductance and anomalous mole fraction behaviour, suggesting the presence of multiple anion binding sites within the channel pore. However, under steady-state conditions SCN– conductance is very low. Here I show, using patch clamp recording from CFTR-transfected mammalian cell lines, that under steady-state conditions neither SCN– conductance nor SCN– permeability show anomalous mole fraction behaviour. Instead, SCN– conductance, permeability, and block of Cl– permeation can all be reproduced by a rate theory model that assumes only a single intrapore anion binding site. These results suggest that under steady-state conditions the interaction between SCN– and the CFTR channel pore can be understood by a simple model whereby SCN– ions enter the pore more easily than Cl–, and bind within the pore more tightly than Cl–. The implications of these findings for investigating and understanding the mechanism of anion permeation are discussed.Key words: chloride channel, permeation, anion binding, multi-ion pore behaviour, rate theory model.