Structure and allosteric effects of low-molecular-weight activators on the protein kinase PDK1
Structure and allosteric effects of low-molecular-weight activators on the protein kinase PDK1
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DOI：
10.1038/nchembio.208
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发表时间：
2009-10-01
影响因子：
14.8
通讯作者：
Biondi, Ricardo M.
中科院分区：
文献类型：
--
作者：
Hindie, Valerie;Stroba, Adriana;Biondi, Ricardo M.
蛋白质磷酸化转导大量细胞内信号。磷酸化介导信号转导的一种机制是促进靶蛋白或相互作用蛋白的构象变化。先前的工作描述了介导 AGC 激酶磷酸化依赖性激活的变构位点。 AGC 激酶 PDK1 通过与底物上的磷酸化基序对接而被激活。在这里，我们展示了与合理开发的低分子量激活剂结合的 PDK1 的晶体学，并使用基于荧光的测定和氘交换实验描述了晶体和溶液中的小化合物诱导的构象变化。我们的结果表明，化合物的结合在靶位点、PIF 结合口袋处产生局部变化，并且在 ATP 结合位点和激活环处产生变构变化。总而言之，我们展示了小化合物诱导的变构变化的分子细节，这些小化合物通过模拟磷酸化依赖性构象变化来触发 PDK1 的激活。
Protein phosphorylation transduces a large set of intracellular signals. One mechanism by which phosphorylation mediates signal transduction is by prompting conformational changes in the target protein or interacting proteins. Previous work described an allosteric site mediating phosphorylation-dependent activation of AGC kinases. The AGC kinase PDK1 is activated by the docking of a phosphorylated motif from substrates. Here we present the crystallography of PDK1 bound to a rationally developed low-molecular-weight activator and describe the conformational changes induced by small compounds in the crystal and in solution using a fluorescence-based assay and deuterium exchange experiments. Our results indicate that the binding of the compound produces local changes at the target site, the PIF binding pocket, and also allosteric changes at the ATP binding site and the activation loop. Altogether, we present molecular details of the allosteric changes induced by small compounds that trigger the activation of PDK1 through mimicry of phosphorylation-dependent conformational changes.