Deconvolution of complex G protein-coupled receptor signaling in live cells using dynamic mass redistribution measurements
Deconvolution of complex G protein-coupled receptor signaling in live cells using dynamic mass redistribution measurements
复制标题
DOI：
10.1038/nbt.1671
复制
发表时间：
2010-09-01
影响因子：
46.9
通讯作者：
Kostenis, Evi
中科院分区：
文献类型：
--
作者：
Schroeder, Ralf;Janssen, Nicole;Kostenis, Evi
基于细胞成分动态质量再分配 (DMR) 的无标记生物传感器技术有望将 GPCR 信号转变成活细胞中实时的复杂光学“指纹”。在这里，我们提出了一种绘制定义无标记反应的细胞机制的策略，并将 DMR 技术与目前 GPCR 药物发现领域最先进的传统第二信使测定进行比较。 DMR 测量的整体性使我们能够 (i) 沿着所有四个 G 蛋白信号通路探测 GPCR 功能，这是目前大多数其他检测平台无法企及的； (ii) 以前所未有的准确性剖析复杂的 GPCR 信号传导模式，甚至在原代人类细胞中也是如此； (iii) 将异源三聚体 G 蛋白定义为复杂光学指纹的触发因素； (iv) 披露之前未检测到的 GPCR 行为特征。我们的结果表明，DMR 技术将对系统生物学和系统药理学以及具有新机制的药物的发现产生重大影响。 (C) 2010 Nature America, Inc. 保留所有权利。
Label-free biosensor technology based on dynamic mass redistribution (DMR) of cellular constituents promises to translate GPCR signaling into complex optical 'fingerprints' in real time in living cells. Here we present a strategy to map cellular mechanisms that define label-free responses, and we compare DMR technology with traditional second-messenger assays that are currently the state of the art in GPCR drug discovery. The holistic nature of DMR measurements enabled us to (i) probe GPCR functionality along all four G-protein signaling pathways, something presently beyond reach of most other assay platforms; (ii) dissect complex GPCR signaling patterns even in primary human cells with unprecedented accuracy; (iii) define heterotrimeric G proteins as triggers for the complex optical fingerprints; and (iv) disclose previously undetected features of GPCR behavior. Our results suggest that DMR technology will have a substantial impact on systems biology and systems pharmacology as well as for the discovery of drugs with novel mechanisms. (C) 2010 Nature America, Inc. All rights reserved.