Laboratory of Molecular Pharmacology, Saint Petersburg State Institute of Technology (Technical University), Moskovskii pr. 26, 190013, Saint Petersburg, Russia.
Laboratory of Regulation of Gene Expression, Institute of Cytology RAS, Tikhoretskii pr. 4, 194064, Saint Petersburg, Russia.
ChemMedChem. 2020 Dec 15;15(24):2521-2529. doi: 10.1002/cmdc.202000579. Epub 2020 Sep 15.
AMP-activated protein kinase (AMPK) is currently the subject of intensive study and active discussions. AMPK performs its functions both at the cellular level, providing the switch between energy-consuming and energy-producing processes, and at the whole body level, particularly, regulating certain aspects of higher nervous activity and behavior. Control of such a 'main switch' compensates dysfunctions and associated diseases. In the present paper, we studied the binding of 3-benzylidene oxindoles to the kinase domain of the AMPK α-subunit, which is thought to prevent its interaction with the autoinhibitory domain and thus result in the AMPK activation. For this purpose, we developed the cellular test system based on the AMPKAR plasmid, which implements the FRET effect, synthesized a number of 3-benzylidene oxindole compounds and simulated their binding to various sites of the kinase domain. The most probable binding site for the studied compounds was established by the correlation of calculated and experimental data. The obtained results allow to analyze various classes of AMPK activators using virtual and high-content screening.
腺苷酸活化蛋白激酶（AMPK）目前是一个备受关注的研究热点，其在细胞水平上发挥着作用，调节着能量消耗和产生的转换，在全身水平上也发挥着作用，特别是调节着高级神经活动和行为的某些方面。对这样一个“主开关”的控制可以补偿功能障碍和相关疾病。在本研究中，我们研究了 3-亚苄基氧吲哚与 AMPKα 亚基激酶结构域的结合，这种结合被认为可以阻止其与自身抑制结构域的相互作用，从而导致 AMPK 的激活。为此，我们开发了基于 AMPKAR 质粒的细胞测试系统，该系统实现了 FRET 效应，合成了一系列 3-亚苄基氧吲哚化合物，并模拟了它们与激酶结构域的各个结合位点的结合。通过计算和实验数据的相关性，确定了研究化合物最可能的结合位点。获得的结果可以使用虚拟和高内涵筛选来分析各种类型的 AMPK 激活剂。
