Uncovering biases in high throughput screens of G-protein coupled receptors.
Uncovering biases in high throughput screens of G-protein coupled receptors.
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DOI：
10.1006/jtbi.2000.2227
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发表时间：
2001-02
影响因子：
2
通讯作者：
P. Woolf;T. Kenakin;J. Linderman
中科院分区：
文献类型：
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作者：
P. Woolf;T. Kenakin;J. Linderman
使用数学模型检查高通量膜结合测定检测G蛋白偶联受体的配体的能力。使用扩展的三元复合物模型开发膜测定模型（Samama等人，1993年，作为基础。通过添加G蛋白活化步骤来模拟配体与全细胞的结合。结果显示，与全细胞中的受体相比，反向激动剂在膜结合测定中与受体的结合更慢且亲和力更低，导致膜测定错过药学上重要的反向激动剂。分析修改，以允许反向激动剂的检测进行了讨论。最后，动力学结合数据显示，以提供有关配体功效的信息。这项工作表明，在检测药物筛选试验中的偏差，并建议技术，以纠正这些偏差的数学建模的效用。
The ability of high throughput membrane binding assays to detect ligands for G-protein coupled receptors was examined using mathematical models. Membrane assay models were developed using the extended ternary complex model (Samama et al., 1993) as a basis. Ligand binding to whole cells was modeled by adding a G-protein activation step. Results show that inverse agonists bind more slowly and with a lower affinity to receptors in the membrane binding assay than to receptors in whole cells, causing the membrane assay to miss pharmaceutically important inverse agonists. Assay modifications to allow detection of inverse agonists are discussed. Finally, kinetic binding data are shown to provide information about ligand efficacy. This work demonstrates the utility of mathematical modeling in detecting biases in drug-screening assay, and also in suggesting techniques to correct those biases.