Department of Oncological Sciences, Department of Pharmacological Sciences, The Tisch Cancer Institute, Mount Sinai Centre for Therapeutic Discovery, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
Department of Oncological Sciences, Department of Pharmacological Sciences, The Tisch Cancer Institute, Mount Sinai Centre for Therapeutic Discovery, Icahn School of Medicine at Mount Sinai, New York, NY, United States.
Methods Enzymol. 2022;667:365-402. doi: 10.1016/bs.mie.2022.03.039. Epub 2022 Apr 25.
Pseudokinases often operate through functionally related enzymes and receptors. A prime example is the pseudokinase KSR (Kinase Suppressor of RAS), which can act as both an amplifier and inhibitor of members in the RAS-MAPK (Mitogen Activated Protein Kinase) signaling pathway. KSR is structurally related to the active RAF kinases over multiple domains; moreover, the pseudokinase domain of KSR forms physical and regulatory complexes with both RAF and MEK through distinct interfaces. Characterization of small molecule interactions on KSR has been used to uncover novel chemical tools and understand the mechanism of action of clinical drugs. Here, we elaborate on assays and structural methods for measuring binding at orthosteric and interfacial binding sites on KSR. These distinct small molecule pockets provide therapeutic paths for targeting KSR1 and KSR2 pseudokinases in disease, including in RAS and RAF mutant cancers.
假激酶通常通过功能相关的酶和受体发挥作用。一个主要的例子是假激酶 KSR（RAS 的激酶抑制剂），它可以作为 RAS-MAPK（丝裂原活化蛋白激酶）信号通路成员的放大器和抑制剂。KSR 在多个结构域上与活性 RAF 激酶具有结构关系；此外，KSR 的假激酶结构域通过不同的界面与 RAF 和 MEK 形成物理和调节复合物。对 KSR 上小分子相互作用的表征已被用于发现新的化学工具，并了解临床药物的作用机制。在这里，我们详细介绍了用于测量 KSR 上正位和界面结合位点结合的测定法和结构方法。这些不同的小分子口袋为靶向 KSR1 和 KSR2 假激酶在疾病中的治疗提供了途径，包括在 RAS 和 RAF 突变型癌症中。
