Department of Chemical Engineering and Biotechnology, University of Applied Sciences Darmstadt, Haardtring 100, 64295 Darmstadt, Germany.
Int J Mol Sci. 2022 Oct 4;23(19):11775. doi: 10.3390/ijms231911775.
HDAC8 is an important target in several indication areas including childhood neuroblastoma. Several isozyme selective inhibitors of HDAC8 with L-shaped structures have been developed. A theoretical study has suggested that methionine 274 (M274) would act as a "switch" that controls a transient binding pocket, which is induced upon binding of L-shaped inhibitors. This hypothesis was experimentally examined in this study. The thermostability and functionality of HDAC8 wildtype and mutant variants with exchanged M274 were analyzed using biophysical methods. Furthermore, the binding kinetics of L-shaped and linear reference inhibitors of these HDAC8 variants were determined in order to elucidate the mode of interaction. Exchange of M274 has considerable impact on enzyme activity, but is not the decisive factor for selective recognition of HDAC8 by L-shaped inhibitors.
组蛋白去乙酰化酶 8（HDAC8）是包括儿童神经母细胞瘤在内的多个适应证领域的重要靶点。已经开发出了几种具有 L 型结构的组蛋白去乙酰化酶 8 同工酶选择性抑制剂。一项理论研究表明，蛋氨酸 274（M274）将充当“开关”，控制一个瞬时结合口袋，该口袋在结合 L 型抑制剂时被诱导。本研究对此假说进行了实验检验。使用生物物理方法分析了野生型和突变型 HDAC8 及其交换 M274 的变体的热稳定性和功能。此外，还确定了这些 HDAC8 变体的 L 型和线性参考抑制剂的结合动力学，以阐明相互作用模式。M274 的交换对酶活性有很大的影响，但不是 L 型抑制剂选择性识别 HDAC8 的决定性因素。
