Solaz-Fuster M Carmen, Gimeno-Alcañiz José Vicente, Casado Marta, Sanz Pascual
Instituto de Biomedicina de Valencia CSIC, Jaime Roig 11, 46010-Valencia, Spain.
Cell Signal. 2006 Oct;18(10):1702-12. doi: 10.1016/j.cellsig.2006.01.021. Epub 2006 Mar 6.
AMP-activated protein kinase (AMPK) is a serine/threonine protein kinase that acts as a sensor of cellular energy charge. Once activated it switches on catabolic pathways and switches off many ATP-consuming processes (anabolic pathways) to preserve the energy status of the cell. In order to identify new targets of AMPK action we have performed a two-hybrid screening of a human pancreas cDNA library. As a result, we have identified TRIP6 as a novel target of AMPK action. This protein belongs to the zyxin family of proteins located at the focal adhesion plaques in the plasma membrane, although they may also travel to the nucleus, where they have regulatory properties. We confirmed the physical interaction between the catalytic subunit (AMPK-alpha2) of the AMPK complex and TRIP6 in mammalian cells by two-hybrid and co-immunoprecipitation assays. We also showed that AMPK was able to phosphorylate in vitro TRIP6 at the N-terminus. Finally, we present evidence that transcriptional co-activator properties of TRIP6 were enhanced by AMPK action.
AMP激活的蛋白激酶（AMPK）是一种丝氨酸/苏氨酸蛋白激酶，作为细胞能量状态的传感器。一旦被激活，它会开启分解代谢途径并关闭许多消耗ATP的过程（合成代谢途径），以维持细胞的能量状态。为了确定AMPK作用的新靶点，我们对人胰腺cDNA文库进行了双杂交筛选。结果，我们确定TRIP6是AMPK作用的一个新靶点。这种蛋白质属于位于质膜粘着斑的zyxin蛋白家族，尽管它们也可能进入细胞核并在那里具有调节特性。我们通过双杂交和免疫共沉淀试验证实了AMPK复合物的催化亚基（AMPK-α2）与TRIP6在哺乳动物细胞中的物理相互作用。我们还表明，AMPK能够在体外使TRIP6的N端磷酸化。最后，我们提供证据表明，AMPK的作用增强了TRIP6的转录共激活特性。
