Inhaled anesthetics were introduced in surgery over a century ago. To this day, the molecular mechanism of anesthetic action remains largely unknown. However, ion-channels of neuronal membranes are believed to be the most- likely molecular targets of inhaled anesthetics. In the study presented here, we investigated the interaction of a simplified ion-channel system, gramicidin, with halothane, a small haloalkane inhaled anesthetic in various environments. Fluorescence-quenching experiments of gramicidin D in dioleoylphosphatidylcholine (DOPC) large unilamellar vesicles (LUVS) have shown that halothane can directly interact with the ion channel (KSV = 66 M–1). Halothane quenched the fluorescence from tryptophan residues located at the lipid bilayer – aqueous interfaces as well as those tryptophans located deeper in the bilayer. Quenching data from gramicidin D in sodium dodecyl sulfide (SDS) micelles revealed that the tryptophan residues located at the micelle–solvent interface were preferentially quenched by halothane (KSV = 22 M–1). In 1-octanol, fluorescence quenching was observed, but with a lower KSV value (KSV = 6 M–1) than in DOPC LUVS and SDS micelles. Taken together, these results indicate that halothane interactions with gramicidin, mediated by a lipid bilayer, are the strongest, and that the mechanism of anesthetic action may also be lipid-mediated.
Determination of the depth of BODIPY probes in model membranes by parallax analysis of fluorescence quenching
