JNK-IN-8 KINOMEscan

The KINOMEscan assay platform is based on a competition binding assay that is run for a compound of interest against each of a panel of 317 to 456 kinases. The assay has three components: a kinase-tagged phage, a test compound, and an immobilized ligand that the compound competes with to displace the kinase. The amount of kinase bound to the immobilized ligand is determined using quantitative PCR of the DNA tag. Results for each kinase are reported as "Percent of control", where the control is DMSO and where a 100% result means no inhibition of kinase binding to the ligand in the presence of the compound, and where low percent results mean strong inhibition. The KINOMEscan data are presented graphically on TREEspot Kinase Dendrograms (http://www.kinomescan.com/Tools---Resources/Study-Reports---Data-Analysis). For this study, HMS LINCS investigators have graphed results for kinases classified as 35 "percent of control" (in the presence of the compound, the kinase is 35% as active for binding ligand in the presence of DMSO), 5 "percent of control" and 1 "percent of control".

KINOMEscan检测平台基于竞争性结合检测技术，针对目标化合物与317至456种激酶组成的检测面板逐一开展检测。该检测包含三个组分：激酶标记噬菌体（kinase-tagged phage）、受试化合物，以及可与受试化合物竞争结合并置换激酶的固定化配体。通过对DNA标签进行定量PCR（quantitative PCR），即可测定与固定化配体结合的激酶总量。每种激酶的检测结果以“对照百分比（Percent of control）”表示，其中对照为二甲基亚砜（DMSO）：当结果为100%时，代表受试化合物存在时激酶与配体的结合未受抑制；百分比数值越低，则说明化合物对激酶结合的抑制作用越强。KINOMEscan检测数据可通过TREEspot激酶树状图（TREEspot Kinase Dendrograms）进行可视化呈现，相关工具可访问网址：http://www.kinomescan.com/Tools---Resources/Study-Reports---Data-Analysis。本研究中，HMS LINCS研究人员对三类激酶的检测结果进行了可视化绘制：分别为35种“对照百分比”（即受试化合物存在时，激酶与配体的结合活性仅为DMSO对照条件下的35%）、5种“对照百分比”以及1种“对照百分比”的激酶。