Identification of Tyrosine Phosphatases That Dephosphorylate the Insulin Receptor
Identification of Tyrosine Phosphatases That Dephosphorylate the Insulin Receptor
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DOI：
10.1074/jbc.275.13.9792
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发表时间：
2000-03
期刊：
影响因子：
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通讯作者：
S. Wälchli;M. Curchod;R. Gobert;S. Arkinstall;R. H. van Huijsduijnen
中科院分区：
文献类型：
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作者：
S. Wälchli;M. Curchod;R. Gobert;S. Arkinstall;R. H. van Huijsduijnen
许多重要的药理受体，包括所有细胞因子受体，通过酪氨酸(自动)磷酸化发出信号，然后通过蛋白酪氨酸磷酸酶(PTPs)的作用恢复到原始状态。确定PTPs对受体底物的特异性对于了解信号是如何调控的以及作为配体模拟物的特定PTP抑制剂的开发都是至关重要的。我们已经建立了一种系统的方法来寻找受体特异性的PTP，并用胰岛素受体激酶验证了这种方法。我们已经在膜结合试验中测试了几乎所有已知的人PTP(45)，使用的是“底物捕捉”PTP突变体。这些结果与二次去磷酸化试验相结合，证实并扩展了早期的发现，即PTP-1b和T细胞PTP是胰岛素受体激酶的生理酶。我们证明，这种方法可以迅速减少以特定受体或其他磷蛋白为底物的PTP的数量。
Many pharmacologically important receptors, including all cytokine receptors, signal via tyrosine (auto)phosphorylation, followed by resetting to their original state through the action of protein tyrosine phosphatases (PTPs). Establishing the specificity of PTPs for receptor substrates is critical both for understanding how signaling is regulated and for the development of specific PTP inhibitors that act as ligand mimetics. We have set up a systematic approach for finding PTPs that are specific for a receptor and have validated this approach with the insulin receptor kinase. We have tested nearly all known human PTPs (45) in a membrane binding assay, using “substrate-trapping” PTP mutants. These results, combined with secondary dephosphorylation tests, confirm and extend earlier findings that PTP-1b and T-cell PTP are physiological enzymes for the insulin receptor kinase. We demonstrate that this approach can rapidly reduce the number of PTPs that have a particular receptor or other phosphoprotein as their substrate.