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  • Stable polymer bilayers for protein channel recordings at high guanidinium chloride concentrations.

Stable polymer bilayers for protein channel recordings at high guanidinium chloride concentrations.

Biophysical journal (2021-02-23)
Luning Yu, Xinqi Kang, Mohammad Amin Alibakhshi, Mikhail Pavlenok, Michael Niederweis, Meni Wanunu
ABSTRACT

The use of chaotropic reagents is common in biophysical characterization of biomolecules. When the study involves transmembrane protein channels, the stability of the protein channel and supporting bilayer membrane must be considered. In this letter, we show that planar bilayers composed of poly(1,2-butadiene)-b-poly(ethylene oxide) diblock copolymer are stable and leak-free at high guanidinium chloride concentrations, in contrast to diphytanoyl phosphatidylcholine bilayers, which exhibit deleterious leakage under similar conditions. Furthermore, insertion and functional analysis of channels such as α-hemolysin and MspA are straightforward in these polymer membranes. Finally, we demonstrate that α-hemolysin channels maintain their structural integrity at 2 M guanidinium chloride concentrations using blunt DNA hairpins as molecular reporters.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
α-Hemolysin from Staphylococcus aureus, lyophilized powder, Protein ~60 % by Lowry, ≥10,000 units/mg protein