Stable polymer bilayers for protein channel recordings at high guanidinium chloride concentrations.
Stable polymer bilayers for protein channel recordings at high guanidinium chloride concentrations.
复制标题
用于高氯化胍浓度下蛋白质通道记录的稳定聚合物双层。
DOI：
10.1016/j.bpj.2021.02.019
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发表时间：
2021
影响因子：
3.4
通讯作者：
Wanunu,Meni
中科院分区：
文献类型：
--
作者：
Yu,Luning;Kang,Xinqi;Alibakhshi,MohammadAmin;Pavlenok,Mikhail;Niederweis,Michael;Wanunu,Meni
离液剂的使用在生物分子的生物物理表征中很常见。当研究涉及跨膜蛋白通道时，必须考虑蛋白通道和支撑双层膜的稳定性。在这封信中，我们表明，由聚（1,2-丁二烯）-b-聚（环氧乙烷）二嵌段共聚物组成的平面双层在高氯化胍浓度下是稳定且无泄漏的，而二植酰磷脂酰胆碱双层在类似条件下表现出有害的泄漏。此外，在这些聚合物膜中，α-溶血素和 MspA 等通道的插入和功能分析非常简单。最后，我们证明 α-溶血素通道在 2 M 氯化胍浓度下使用钝 DNA 发夹作为分子报告基因保持其结构完整性。
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 asα-hemolysin and MspA are straightforward in these polymer membranes. Finally, we demonstrate thatα-hemolysin channels maintain their structural integrity at 2 M guanidinium chloride concentrations using blunt DNA hairpins as molecular reporters.