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用于高盐酸胍浓度下蛋白质通道记录的稳定聚合物双层膜。

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

机构信息

Department of Physics.

Department of Bioengineering.

出版信息

Biophys J. 2021 May 4;120(9):1537-1541. doi: 10.1016/j.bpj.2021.02.019. Epub 2021 Feb 20.

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.

摘要

在生物分子的生物物理特性描述中,离液试剂的使用很常见。当研究涉及跨膜蛋白通道时,必须考虑蛋白质通道和支撑双层膜的稳定性。在这封信中,我们表明,与二棕榈酰磷脂酰胆碱双层膜相比,由聚(1,2-丁二烯)-b-聚(氧化乙烯)嵌段共聚物组成的平面双层膜在高浓度胍盐下稳定且无泄漏,在相似条件下,二棕榈酰磷脂酰胆碱双层膜会表现出有害的泄漏。此外,α-溶血素和 MspA 等通道的插入和功能分析在这些聚合物膜中很简单。最后,我们使用钝状 DNA 发夹作为分子报告物证明,α-溶血素通道在 2 M 胍盐浓度下保持其结构完整性。

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