Proteomic screen finds pSer/pThr-binding domain localizing Plk1 to mitotic substrates
Proteomic screen finds pSer/pThr-binding domain localizing Plk1 to mitotic substrates
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DOI：
10.1126/science.1079079
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发表时间：
2003-02-21
期刊：
影响因子：
56.9
通讯作者：
Yaffe, MB
中科院分区：
文献类型：
--
作者：
Elia, AEH;Cantley, LC;Yaffe, MB
我们已经开发了一种蛋白质组学的方法来确定磷酸肽结合域，调节激酶依赖的信号通路。偏向于特定蛋白激酶磷酸化基序的部分简并磷酸肽的固定化文库用于分离与被该激酶磷酸化的蛋白质结合的磷酸结合结构域。将这种方法应用于细胞周期蛋白依赖性激酶（Cdks），我们确定了有丝分裂激酶polo样激酶1（Plk 1）的polo盒结构域（PBD）作为一个特定的磷酸丝氨酸（pSer）或磷酸苏氨酸（pThr）结合结构域，并确定了其最佳结合基序。该基序存在于已知的Plk 1底物如Cdc 25中，并且含有该基序的最佳磷酸肽破坏PBD-底物结合和PBD向中心体的定位。这一发现揭示了Plk 1如何定位于细胞内的特定位点，以响应Cdk在这些位点的磷酸化，并提供了一种将Plk 1激酶结构域靶向其底物的结构机制。
We have developed a proteomic approach for identifying phosphopeptide binding domains that modulate kinase-dependent signaling pathways. An immobilized library of partially degenerate phosphopeptides biased toward a particular protein kinase phosphorylation motif is used to isolate phospho-binding domains that bind to proteins phosphorylated by that kinase. Applying this approach to cyclin-dependent kinases (Cdks), we identified the polo-box domain (PBD) of the mitotic kinase polo-like kinase 1 (Plk1) as a specific phosphoserine (pSer) or phosphothreonine (pThr) binding domain and determined its optimal binding motif. This motif is present in known Plk1 substrates such as Cdc25, and an optimal phosphopeptide containing the motif disrupted PBD-substrate binding and localization of the PBD to centrosomes. This finding reveals how Plk1 can localize to specific sites within cells in response to Cdk phosphorylation at those sites and provides a structural mechanism for targeting the Plk1 kinase domain to its substrates.