Interfacing lipid bilayer nanodiscs and silicon photonic sensor arrays for multiplexed protein-lipid and protein-membrane protein interaction screening.
Interfacing lipid bilayer nanodiscs and silicon photonic sensor arrays for multiplexed protein-lipid and protein-membrane protein interaction screening.
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DOI：
10.1021/ac3037359
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发表时间：
2013-03-05
影响因子：
7.4
通讯作者：
Bailey, Ryan C.
中科院分区：
文献类型：
--
作者：
Sloan, Courtney D. Kuhnline;Marty, Michael T.;Sligar, Stephen G.;Bailey, Ryan C.
可溶性蛋白是许多生物化学信号通路的关键介质，通过脂质双分子层和膜结合受体直接相互作用。细胞膜组分参与许多重要的生物过程，包括病毒感染、血液凝固和信号转导，因此，它们是治疗剂的共同靶点。因此，研究细胞膜相互作用的分析方法的发展是至关重要的。在此，我们整合了两项关键技术，硅光子微环谐振器阵列和磷脂双层纳米盘，它们一起允许多路筛选与脂质和膜嵌入靶标的可溶性蛋白相互作用。微环谐振器阵列是一种本质上可复用的，无标记的分析平台，以前已应用于研究蛋白质-蛋白质，蛋白质-核酸和核酸-核酸相互作用。纳米圆盘是蛋白质稳定的脂质组件，代表了一种方便的结构来模拟天然磷脂双分子层，研究膜组成的影响，并溶解膜嵌入的目标。利用纳米圆盘支持的脂质双层对氧化钝化硅的天然亲和力，我们通过直接物理吸附组装了单和多传感器阵列，表征了静电对纳米圆盘附着的影响。利用模型系统，我们证明了该平台在平行筛选蛋白质与纳米圆盘嵌入的脂质、糖脂质和膜蛋白相互作用方面的适用性。
Soluble proteins are key mediators of many biochemical signaling pathways via direct interaction with the lipid bilayer and via membrane-bound receptors. Components of the cell membrane are involved in many important biological processes, including viral infection, blood clotting, and signal transduction, and as such, they are common targets of therapeutic agents. Therefore, the development of analytical approaches to study interactions at the cell membrane is of critical importance. Herein, we integrate two key technologies, silicon photonic microring resonator arrays and phospholipid bilayer nanodiscs, which together allow multiplexed screening of soluble protein interactions with lipid and membrane-embedded targets. Microring resonator arrays are an intrinsically multiplexable, label-free analysis platform that has previously been applied to studying protein-protein, protein-nucleic acid, and nucleic acid-nucleic acid interactions. Nanodiscs are protein-stabilized lipid assemblies that represent a convenient construct to mimic the native phospholipid bilayer, investigate the effects of membrane composition, and solubilize membrane-embedded targets. Exploiting the natural affinity of nanodisc-supported lipid bilayers for oxide-passivated silicon, we assembled single and multiplex sensor arrays via direct physisorption, characterizing electrostatic effects on nanodisc attachment. Using model systems, we demonstrate the applicability of this platform for the parallel screening of protein interactions with nanodisc-embedded lipids, glycolipids, and membrane proteins.
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影响因子：
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