Allosteric activation of the protein kinase PDK1 with low molecular weight compounds
Allosteric activation of the protein kinase PDK1 with low molecular weight compounds
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DOI：
10.1038/sj.emboj.7601416
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发表时间：
2006-11-29
期刊：
影响因子：
11.4
通讯作者：
Biondi, Ricardo M.
中科院分区：
文献类型：
--
作者：
Engel, Matthias;Hindie, Valerie;Biondi, Ricardo M.
生物体在很大程度上依赖于蛋白质磷酸化来传递细胞内信号。蛋白质的磷酸化通常会引起构象变化，这是引发下游细胞事件的原因。蛋白激酶本身经常受到磷酸化的调节。最近，我们和其他人提出了疏水基序(HM)的磷酸化调节AGC蛋白激酶组中许多成员的构象和活性的分子机制。在这里，我们开发了特定的低分子化合物，这些化合物以HM/PIF-Pocket为靶标，能够通过调节磷酸化依赖的构象转换来变构激活磷酸肌醇依赖的蛋白激酶1(PDK1)。通过对PDK1的诱变、化合物类似物的合成、相互作用-置换研究和等温滴定量热实验，对这些化合物的作用机理进行了表征。我们的结果提高了通过一种新的作用模式开发针对AGC激酶的药物的可能性，并可能激励未来合理开发能够调节其他蛋白质中依赖于磷酸化的构象转变的化合物。
Organisms rely heavily on protein phosphorylation to transduce intracellular signals. The phosphorylation of a protein often induces conformational changes, which are responsible for triggering downstream cellular events. Protein kinases are themselves frequently regulated by phosphorylation. Recently, we and others proposed the molecular mechanism by which phosphorylation at a hydrophobic motif (HM) regulates the conformation and activity of many members of the AGC group of protein kinases. Here we have developed specific, low molecular weight compounds, which target the HM/PIF-pocket and have the ability to allosterically activate phosphoinositide-dependent protein kinase 1 (PDK1) by modulating the phosphorylation-dependent conformational transition. The mechanism of action of these compounds was characterized by mutagenesis of PDK1, synthesis of compound analogs, interaction-displacement studies and isothermal titration calorimetry experiments. Our results raise the possibility of developing drugs that target the AGC kinases via a novel mode of action and may inspire future rational development of compounds with the ability to modulate phosphorylation-dependent conformational transitions in other proteins.