Abstract
This paper describes the use of a newly-developed micro-chip bilayer platform to examine the electrophysiological properties of the prokaryotic voltage-gated sodium channel pore (NavSp) from Silicibacter pomeroyi. The platform allows up to 6 bilayers to be analysed simultaneously. Proteoliposomes were incorporated into suspended lipid bilayers formed within the microfluidic bilayer chips. The chips provide access to bilayers from either side, enabling the fast and controlled titration of compounds. Dose-dependent modulation of the opening probability by the channel blocking drug nifedipine was measured and its IC50 determined.
Highlights
Human voltage-gated sodium channels (Nav's) are large pseudotetrameric integral membrane proteins which play important roles in many physiological processes and are linked to channelopathies including epilepsy, pain disorders and cardiac conditions such as long QT syndrome and Brugada syndrome [1,2,3]
The electrical behaviour is generally similar to data from Shaya et al [5] for the NavSp1 pore-only construct, inserted into giant unilamellar vesicles (GUVs) made of 1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) for planar patch clamp electrophysiology
The properties of the NavSp pore were characterised using a microfluidic platform that records from 6 bilayers simultaneously
Summary
Human voltage-gated sodium channels (Nav's) are large pseudotetrameric integral membrane proteins which play important roles in many physiological processes and are linked to channelopathies including epilepsy, pain disorders and cardiac conditions such as long QT syndrome and Brugada syndrome [1,2,3]. They are major targets for drug development, and are of considerable interest to the pharmaceutical industry. We have expressed the isolated tetrameric pore-only construct of the prokaryotic voltage-gated sodium channel NavSp from Silicibacter pomeroyi. This pore-only construct has previously been shown to be correctly folded, more thermally stable than the full length channel and PLOS ONE | DOI:10.1371/journal.pone.0131286 July 6, 2015
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