Surface plasmon resonance for high-throughput ligand screening of membrane-bound proteins.
Surface plasmon resonance for high-throughput ligand screening of membrane-bound proteins.
复制标题
DOI：
10.1002/biot.200900195
复制
发表时间：
2009-11
影响因子：
4.7
通讯作者：
Oh, Sang-Hyun
中科院分区：
文献类型：
--
作者：
Maynard, Jennifer A;Lindquist, Nathan C;Sutherland, Jamie N;Lesuffleur, Antoine;Warrington, Arthur E;Rodriguez, Moses;Oh, Sang-Hyun
基于表面等离子体共振（SPR）的技术允许快速、无标记地表征蛋白质-蛋白质和蛋白质-小分子相互作用，从结合动力学和热力学的定量测量以及复杂样品中的浓度到表位分析。SPR已成为工业和学术环境中的金标准，其中通常详细表征一对可溶性结合配偶体之间的相互作用或筛选分子库以结合单个可溶性蛋白质。尽管取得了这些成功，但该技术才刚刚开始适应膜结合蛋白的需求。包括G蛋白偶联受体（GPCR）、离子通道和其他生长、免疫和细胞受体，这些蛋白质难以原位研究，但代表了药物和生物标志物开发的有希望的靶点。现有的技术，如BIAcore™，已经通过在现有芯片上构建支持的脂质层或囊泡捕获而适用于膜蛋白分析。仍在开发中的新技术将使膜蛋白以天然或接近天然的形式呈现。这些包括SPR纳米孔阵列，其中含有膜蛋白的脂质双层稳定地跨越可从双层两侧寻址的小孔。在这里，我们讨论了目前SPR仪器的成功和SPR纳米孔阵列的潜力，使定量，高通量筛选GPCR配体，生物标志物的发现涉及膜结合蛋白和基本的细胞生物学。
Technology based on surface plasmon resonance (SPR) has allowed rapid, label-free characterization of protein-protein and protein-small molecule interactions, from quantitative measurements of binding kinetics and thermodynamics and concentrations in complex samples to epitope analysis. SPR has become the gold standard in industrial and academic settings, in which typically the interaction between a pair of soluble binding partners is characterized in detail or a library of molecules is screened for binding against a single soluble protein. In spite of these successes, the technology is only beginning to be adapted to the needs of membrane-bound proteins. Including G protein-coupled receptors (GPCR), ion channels and other growth, immune and cellular receptors, these proteins are difficult to study in situ but represent promising targets for drug and biomarker development. Existing technologies, such as BIAcore™, have been adapted for membrane protein analysis by building supported lipid layers or vesicle capture on existing chips. Newer technologies, still in development, will allow membrane proteins to be presented in native or near-native formats. These include SPR nanopore arrays, in which lipid bilayers containing membrane proteins stably span small pores that are addressable from both sides of the bilayer. Here, we discuss successes with current SPR instrumentation and the potential for SPR nanopore arrays to enable quantitative, high-throughput screening of GPCR ligands, biomarker discovery involving membrane bound proteins and basic cellular biology.