Self-Assembly of Mammalian-Cell Membranes on Bioelectronic Devices with Functional Transmembrane Proteins
Self-Assembly of Mammalian-Cell Membranes on Bioelectronic Devices with Functional Transmembrane Proteins
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DOI：
10.1021/acs.langmuir.0c00804
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发表时间：
2020-07-07
期刊：
影响因子：
3.9
通讯作者：
Daniel, Susan
中科院分区：
文献类型：
--
作者：
Liu, Han-Yuan;Pappa, Anna-Maria;Daniel, Susan
跨膜蛋白 (TMP) 调节细胞表面发生的过程，是跨膜信息流的重要守门人。鉴于膜的复杂环境及其对蛋白质构象、迁移性、生物分子相互作用和活性的影响，TMP 很难研究。我们首次创建了支撑在透明导电聚合物表面上的哺乳动物生物膜，这使得能够在其天然膜环境中对 TMP 功能进行双重电学和光学监测。含有 ATP 门控 P2X2 离子通道的哺乳动物质膜囊泡在生物相容性聚合物垫上自组装，可在 ATP 暴露期间转导离子通量的变化。该平台保持了天然质膜的复杂性、其成分的流动性以及对离子通道功能至关重要的蛋白质方向。我们展示了使用显微镜的双模态读数，通过单粒子跟踪和使用电阻抗光谱检测 P2X2 的 ATP 门控来表征蛋白质迁移率。 TMP 活性的测量对于疼痛感知、神经活动和感觉活动非常重要，为药物筛选和生物传感应用带来了新的可能性。
Transmembrane proteins (TMPs) regulate processes occurring at the cell surface and are essential gatekeepers of information flow across the membrane. TMPs are difficult to study, given the complex environment of the membrane and its influence on protein conformation, mobility, biomolecule interaction, and activity. For the first time, we create mammalian biomembranes supported on a transparent, electrically conducting polymer surface, which enables dual electrical and optical monitoring of TMP function in its native membrane environment. Mammalian plasma membrane vesicles containing ATP-gated P2X2 ion channels self-assemble on a biocompatible polymer cushion that transduces the changes in ion flux during ATP exposure. This platform maintains the complexity of the native plasma membrane, the fluidity of its constituents, and protein orientation critical to ion channel function. We demonstrate the dual-modality readout using microscopy to characterize protein mobility by single-particle tracking and sensing of ATP gating of P2X2 using electrical impedance spectroscopy. This measurement of TMP activity important for pain sensing, neurological activity, and sensory activity raises new possibilities for drug screening and biosensing applications.