Optical waveguide lightmode spectroscopic techniques for investigating membrane-bound ion channel activities.

Székács, Inna and Kaszás, Nóra and Gróf, Pál and Erdélyi, Katalin and Szendrő, István and Mihalik, Balázs and Pataki, Ágnes and Antoni, Ferenc A. and Madarász, Emilia (2013) Optical waveguide lightmode spectroscopic techniques for investigating membrane-bound ion channel activities. PLOS ONE, 8 (12). pp. 1-11. ISSN 1932-6203

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Abstract

Optical waveguide lightmode spectroscopic (OWLS) techniques were probed for monitoring ion permeation through channels incorporated into artificial lipid environment. A novel sensor set-up was developed by depositing liposomes or cell-derived membrane fragments onto hydrophilic polytetrafluoroethylene (PTFE) membrane. The fibrous material of PTFE membrane could entrap lipoid vesicles and the water-filled pores provided environment for the hydrophilic domains of lipid-embedded proteins. The sensor surface was kept clean from the lipid holder PTFE membrane by a water- and ion-permeable polyethylene terephthalate (PET) mesh. The sensor set-up was tested with egg yolk lecithin liposomes containing gramicidin ion channels and with cell-derived membrane fragments enriched in GABA-gated anion channels. The method allowed monitoring the move of Na(+) and organic cations through gramicidin channels and detecting the Cl(-)-channel functions of the (alpha5beta2gamma2) GABAA receptor in the presence or absence of GABA and the competitive GABA-blocker bicuculline.

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