Bhat Ratan, Xue Yafeng, Berg Stefan, Hellberg Sven, Ormö Mats, Nilsson Yvonne, Radesäter Ann-Cathrin, Jerning Eva, Markgren Per-Olof, Borgegård Thomas, Nylöf Martin, Giménez-Cassina Alfredo, Hernández Félix, Lucas Jose J, Díaz-Nido Javier, Avila Jesús
AstraZeneca R&D, 15185 Södertälje, Sweden, AstraZeneca R&D, 43183 Mölndal, Sweden.
J Biol Chem. 2003 Nov 14;278(46):45937-45. doi: 10.1074/jbc.M306268200. Epub 2003 Aug 19.
Glycogen synthase kinase 3 (GSK3) is a serine/threonine kinase that has been implicated in pathological conditions such as diabetes and Alzheimer's disease. We report the characterization of a GSK3 inhibitor, AR-A014418, which inhibits GSK3 (IC50 = 104 +/- 27 nM), in an ATP-competitive manner (Ki = 38 nM). AR-A014418 does not significantly inhibit cdk2 or cdk5 (IC50 > 100 microM) or 26 other kinases demonstrating high specificity for GSK3. We report the co-crystallization of AR-A014418 with the GSK3beta protein and provide a description of the interactions within the ATP pocket, as well as an understanding of the structural basis for the selectivity of AR-A014418. AR-A014418 inhibits tau phosphorylation at a GSK3-specific site (Ser-396) in cells stably expressing human four-repeat tau protein. AR-A014418 protects N2A neuroblastoma cells against cell death mediated by inhibition of the phosphatidylinositol 3-kinase/protein kinase B survival pathway. Furthermore, AR-A014418 inhibits neurodegeneration mediated by beta-amyloid peptide in hippocampal slices. AR-A014418 may thus have important applications as a tool to elucidate the role of GSK3 in cellular signaling and possibly in Alzheimer's disease. AR-A014418 is the first compound of a family of specific inhibitors of GSK3 that does not significantly inhibit closely related kinases such as cdk2 or cdk5.
糖原合酶激酶3（GSK3）是一种丝氨酸/苏氨酸激酶，与糖尿病和阿尔茨海默病等病理状况有关。我们报告了一种GSK3抑制剂AR - A014418的特性，它以ATP竞争性方式抑制GSK3（IC50 = 104±27 nM），抑制常数（Ki = 38 nM）。AR - A014418对细胞周期蛋白依赖性激酶2（cdk2）或细胞周期蛋白依赖性激酶5（cdk5）无显著抑制作用（IC50>100μM），对其他26种激酶也无明显抑制，显示出对GSK3的高度特异性。我们报道了AR - A014418与GSK3β蛋白的共结晶，并描述了ATP口袋内的相互作用，以及对AR - A014418选择性的结构基础的理解。AR - A014418在稳定表达人四重复tau蛋白的细胞中，抑制GSK3特异性位点（Ser - 396）的tau蛋白磷酸化。AR - A014418保护N2A神经母细胞瘤细胞免受因磷脂酰肌醇3激酶/蛋白激酶B生存途径抑制介导的细胞死亡。此外，AR - A014418抑制海马切片中β淀粉样肽介导的神经退行性变。因此，AR - A014418作为一种工具，在阐明GSK3在细胞信号传导以及可能在阿尔茨海默病中的作用方面可能具有重要应用。AR - A014418是GSK3特异性抑制剂家族中的首个化合物，对密切相关的激酶如cdk2或cdk5无显著抑制作用。
