Rapid microfluidic perfusion enabling kinetic studies of lipid ion channels in a bilayer lipid membrane chip.
Rapid microfluidic perfusion enabling kinetic studies of lipid ion channels in a bilayer lipid membrane chip.
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快速微流体灌注能够对双层脂质膜芯片中的脂质离子通道进行动力学研究。
DOI：
10.1007/s10439-011-0323-4
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发表时间：
2011
影响因子：
3.8
通讯作者：
Devoe,DonL
中科院分区：
文献类型：
--
作者：
Shao,Chenren;Sun,Bing;Colombini,Marco;Devoe,DonL
越来越多的人认识到脂质在离子通道生理学中起着关键作用，无论是通过脂质离子通道的动态形成和溶解，还是通过间接调节蛋白质离子通道。由于现有技术不能快速调节离子通道研究中使用的人工双层脂质膜的局部（生物）化学条件，阐明这些脂质-脂质和脂质-蛋白相互作用的动力学的能力受到限制。在这里，我们展示了一种微流体系统，它支持将试剂异常快速地灌注到芯片上的双层脂质膜上，从而可以探测脂质离子通道对膜边界条件动态变化的响应。热塑性微流控系统可以在不到1 s的时间内将试剂初始灌注到膜上，并可以直接测量时间常数低于10 s的动力学行为。该平台应用于神经酰胺的动力学研究，神经酰胺是一种生物学上重要的脂质，已知可以自组装成跨膜离子通道，以响应小离子（La3+）和蛋白质（Bcl-xLmutant）的动态处理。这些结果揭示了膜生物物理学研究技术的广阔潜力，包括脂质离子通道动力学，脂质-蛋白质相互作用，以及脂质微结构域对蛋白质离子通道的调节。
There is growing recognition that lipids play key roles in ion channel physiology, both through the dynamic formation and dissolution of lipid ion channels and by indirect regulation of protein ion channels. Because existing technologies cannot rapidly modulate the local (bio)chemical conditions at artificial bilayer lipid membranes used in ion channel studies, the ability to elucidate the dynamics of these lipid–lipid and lipid–protein interactions has been limited. Here we demonstrate a microfluidic system supporting exceptionally rapid perfusion of reagents to an on-chip bilayer lipid membrane, enabling the responses of lipid ion channels to dynamic changes in membrane boundary conditions to be probed. The thermoplastic microfluidic system allows initial perfusion of reagents to the membrane in less than 1 s, and enables kinetic behaviors with time constants below 10 s to be directly measured. Application of the platform is demonstrated toward kinetic studies of ceramide, a biologically important lipid known to self-assemble into transmembrane ion channels, in response to dynamic treatments of small ions (La3+) and proteins (Bcl-xLmutant). The results reveal the broader potential of the technology for studies of membrane biophysics, including lipid ion channel dynamics, lipid–protein interactions, and the regulation of protein ion channels by lipid micro domains.