利用受体蛋白酪氨酸磷酸酶（RPTP）小干扰RNA（siRNA）-受体酪氨酸激酶（RTK）底物筛选，鉴定RPTP作为RTK调节剂。
Identification of receptor protein tyrosine phosphatases (RPTPs) as regulators of receptor tyrosine kinases (RTKs) using an RPTP siRNA-RTK substrate screen.
作者信息
Lee Hojin, Bennett Anton M
机构信息
Department of Pharmacology, Yale University School of Medicine, SHM B226D, 333 Cedar Street, New Haven, CT, 06520-8066, USA.
出版信息
Methods Mol Biol. 2015;1233:111-20. doi: 10.1007/978-1-4939-1789-1_11.
Receptor tyrosine kinase (RTK) signaling exists in equilibrium between RTK tyrosyl phosphorylation and RTK tyrosyl dephosphorylation. Despite a detailed understanding of RTK tyrosyl phosphorylation, much less is known about RTK tyrosyl dephosphorylation. The receptor PTPs (RPTPs) are outstanding targets for the dephosphorylation of RTKs because of their mutual membrane proximity. In this chapter, we describe how to identify RPTPs that modulate the activity of RTKs using a siRNA screen and commercially available proteomic applications. The validation of putative RTKs as RPTP substrates using substrate-trapping approaches is detailed.
受体酪氨酸激酶（RTK）信号传导存在于RTK酪氨酸磷酸化和RTK酪氨酸去磷酸化之间的平衡状态。尽管对RTK酪氨酸磷酸化有了详细的了解，但对RTK酪氨酸去磷酸化的了解却少得多。受体蛋白酪氨酸磷酸酶（RPTPs）因其与膜的相互接近性而成为RTK去磷酸化的重要靶点。在本章中，我们描述了如何使用siRNA筛选和市售的蛋白质组学应用来鉴定调节RTK活性的RPTPs。详细介绍了使用底物捕获方法验证假定的RTK作为RPTP底物的过程。
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