Institute of Pharmaceutical Chemistry, Goethe-University Frankfurt, 60438 Frankfurt, Germany.
Structural Genomics Consortium, BMLS, Goethe-University Frankfurt, 60438 Frankfurt, Germany.
STAR Protoc. 2021 Aug 5;2(3):100717. doi: 10.1016/j.xpro.2021.100717. eCollection 2021 Sep 17.
Typical enzymatic inhibition assays often demonstrate improved potency for kinase covalent inhibitors compared to reversible inhibitors. This can primarily be attributed to the irreversible mode of action and could affect the evaluations of the ATP-competitive nature of covalent inhibitors, hindering optimization of these compounds. Here, we describe a version of ADP-Glo assay, in which modification of inhibitor incubation time in the presence or absence of ATP enables a quick assessment of relative reversible and irreversible effects of kinase covalent inhibitors. For complete details on the use and execution of this protocol, please refer to Schröder et al. (2020).
典型的酶抑制分析通常显示激酶共价抑制剂比可逆抑制剂具有更高的效力。这主要归因于不可逆作用模式，可能会影响对共价抑制剂的 ATP 竞争性质的评估，从而阻碍这些化合物的优化。在这里，我们描述了一种 ADP-Glo 分析方法，其中抑制剂在存在或不存在 ATP 的情况下孵育时间的改变可快速评估激酶共价抑制剂的相对可逆和不可逆作用。有关该方案的使用和执行的完整详细信息，请参见 Schröder 等人。（2020 年）。
