A Luminescent Biosensor with Increased Dynamic Range for Intracellular cAMP
A Luminescent Biosensor with Increased Dynamic Range for Intracellular cAMP
复制标题
DOI：
10.1021/cb200248h
复制
发表时间：
2011-11-01
影响因子：
4
通讯作者：
Wood, Keith V.
中科院分区：
文献类型：
--
作者：
Binkowski, Brock F.;Butler, Braeden L.;Wood, Keith V.
第二个信使阵营是一个关键的调解人。G蛋白偶联受体激活后的信号转导。G蛋白偶联受体的研究将受益于第二种信使试验，它允许在广泛的动态范围内连续监测cAMP浓度的动态变化。为了实现这一点，我们已经进化出一种用于cAMP的发光生物传感器，以更好地包含生理浓度。存在于活细胞中的范围。与免疫分析相比，进化的生物传感器结构能够准确地跟踪cAMP变化的幅度和动力学，使用的劳动强度要低得多。我们证明了这种结构的实用性，以检测广泛的受体活性，并显示适合用于高通量筛选。
The second messenger cAMP is a key mediator of. signal transduction following activation of G-protein coupled receptors., Investigations on Gs-coupled receptors Would benefit from a second Messenger assay that allows continuous Monitoring of kinetic changes in cAMP concentration over a broad dynamic range. To accomplish this, we have evolved a luminescent biosensor for cAMP to better encompass the physiological Concentration. ranges present in living cells. When compared to an immunoassay, the evolved biosensor construct was able to accurately track both the magnitude and kinetics of cAMP change using a far less labor intensive format. We demonstrate the utility of this construct to detect a broad range of receptor activity, together with showing suitability for use in high throughput screening.