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1. 发明申请

WO2006050257A3 DETECTION OF ION CHANNEL OR RECEPTOR ACTIVITY 审中-公开
标题翻译：检测离子通道或受体活性
公开(公告)号：WO2006050257A3
公开(公告)日：2009-05-07
申请号：PCT/US2005039260
申请日：2005-10-31
申请人： MASSACHUSETTS INST OF TECCHNOL , CHILDRENS MEDICAL CENTER , MARINI DAVIDE M , DESAI BIMAL , DELLING MARKUS , SOLIS DANIEL J , BELCHER ANGELA M , CLAPHAM DAVID E , FEBVAY SEBASTIEN , CARTER BRETT
发明人： MARINI DAVIDE M , DESAI BIMAL , DELLING MARKUS , SOLIS DANIEL J , BELCHER ANGELA M , CLAPHAM DAVID E , FEBVAY SEBASTIEN , CARTER BRETT
IPC分类号： G01N33/551
CPC分类号： C12Q1/005 , B82Y5/00 , B82Y10/00 , G01N21/554 , G01N33/542 , G01N33/54373 , G01N33/6872 , G01N2333/726
摘要： The invention provides nanosensors and nanosensor components for the detection of ion channel activity, receptor activity, or protein protein interactions. Certain of the nanosensor components comprise a nanoparticle and recognition domain. Following contact with cells and, optionally, internalization of the nanosensor component by a cell, the recognition domain binds to a target domain, e.g., a heterologous target domain, of a polypeptide of interest such as an ion channel subunit, G protein coupled receptor (GPCR), or G protein subunit. Ion channel activity, GPCR activity, or altered protein interaction results in a detectable signal. The nanoparticles may be functionalized so that they respond to the presence of an ion by altering their proximity. Certain of the nanosensors utilize the phenomenon of plasmon resonance to produce a signal while others utilize magnetic properties, RET, and/or ion-sensitive moieties. Also provided are polypeptides, e.g., ion channel subunits, comprising a heterologous target domain, and cell lines that express the polypeptides. Further provided are a variety of methods for detecting ion channel activity, receptor activity, or protein interaction and for identifying compounds that modulate one or more of these. In certain embodiments the invention allows the user to detect the activity of specific ion channels even in the presence of other channels that permit passage of the same ion(s) or result in activation of the same downstream targets, thereby achieving improved specificity in high throughput screens while at the same time providing a high signal to noise ratio.
摘要翻译：本发明提供用于检测离子通道活性，受体活性或蛋白质蛋白相互作用的纳米传感器和纳米传感器组分。某些纳米传感器组件包含纳米颗粒和识别结构域。在与细胞接触并且任选地，通过细胞使纳米传感器组分内化后，识别结构域结合感兴趣的多肽的靶结构域例如异源靶结构域，例如离子通道亚基，G蛋白偶联受体（ GPCR）或G蛋白亚基。离子通道活性，GPCR活性或改变的蛋白质相互作用导致可检测的信号。纳米颗粒可以被官能化，使得它们通过改变它们的接近度来响应离子的存在。某些纳米传感器利用等离子体共振现象来产生信号，而另一些利用磁性，RET和/或离子敏感部分。还提供了多肽，例如包含异源靶域的离子通道亚单位，以及表达多肽的细胞系。进一步提供了用于检测离子通道活性，受体活性或蛋白质相互作用以及鉴定调节这些中的一种或多种的化合物的各种方法。在某些实施方案中，本发明允许用户即使在允许相同离子通过或导致相同下游靶的活化的其它通道的存在下也检测特定离子通道的活性，从而实现高通量的改善的特异性屏幕同时提供高信噪比。

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