Department of Chemistry, Lehigh University, Bethlehem, Pennsylvania 18015.
Department of Chemical Engineering, University of Virginia, Charlottesville, Virginia 22903.
J Biol Chem. 2019 Dec 6;294(49):18796-18806. doi: 10.1074/jbc.RA119.010229. Epub 2019 Nov 1.
Receptor protein tyrosine phosphatases (RPTPs) play critical regulatory roles in mammalian signal transduction. However, the structural basis for the regulation of their catalytic activity is not fully understood, and RPTPs are generally not therapeutically targetable. This knowledge gap is partially due to the lack of known natural ligands or selective agonists of RPTPs. Contrary to what is known from structure-function studies of receptor tyrosine kinases (RTKs), RPTP activities have been reported to be suppressed by dimerization, which may prevent RPTPs from accessing their RTK substrates. We report here that homodimerization of protein tyrosine phosphatase receptor J (PTPRJ, also known as DEP-1) is regulated by specific transmembrane (TM) residues. We found that disrupting these interactions destabilizes homodimerization of full-length PTPRJ in cells, reduces the phosphorylation of the known PTPRJ substrate epidermal growth factor receptor (EGFR) and of other downstream signaling effectors, antagonizes EGFR-driven cell phenotypes, and promotes substrate access. We demonstrate these observations in human cancer cells using mutational studies and identified a peptide that binds to the PTPRJ TM domain and represents the first example of an allosteric agonist of RPTPs. The results of our study provide fundamental structural and functional insights into how PTPRJ activity is tuned by TM interactions in cells. Our findings also open up opportunities for developing peptide-based agents that could be used as tools to probe RPTPs' signaling mechanisms or to manage cancers driven by RTK signaling.
受体蛋白酪氨酸磷酸酶 (RPTPs) 在哺乳动物信号转导中发挥着关键的调节作用。然而,其催化活性调节的结构基础尚未完全理解,而且 RPTPs 通常不是治疗的靶向目标。这一知识空白部分归因于缺乏已知的 RPTP 天然配体或选择性激动剂。与受体酪氨酸激酶 (RTKs) 的结构功能研究所表明的情况相反,据报道,RPTP 的活性受到二聚化的抑制,这可能阻止 RPTP 与它们的 RTK 底物接触。我们在此报告,蛋白酪氨酸磷酸酶受体 J(PTPRJ,也称为 DEP-1)的同源二聚化受到特定跨膜 (TM) 残基的调节。我们发现,破坏这些相互作用会使全长 PTPRJ 在细胞中的同源二聚化不稳定,降低已知的 PTPRJ 底物表皮生长因子受体 (EGFR) 和其他下游信号效应物的磷酸化,拮抗 EGFR 驱动的细胞表型,并促进底物接近。我们使用突变研究在人类癌细胞中证明了这些观察结果,并鉴定出一种与 PTPRJ TM 结构域结合的肽,这代表了 RPTPs 的第一个变构激动剂的例子。我们的研究结果为 PTPRJ 活性如何通过细胞中的 TM 相互作用进行调节提供了基本的结构和功能见解。我们的发现还为开发基于肽的药物提供了机会,这些药物可用作研究 RPTPs 信号机制的工具,或用于管理由 RTK 信号驱动的癌症。