Abstract：
Receptor tyrosine kinases (RTKs) are transmembrane receptors activated by a wide diversity of growth factors, cytokines or hormones. They ensure multiple roles in cellular processes, including proliferation, differentiation and survival. They are also crucial drivers of development and progression of multiple cancer types, and represent important drug targets. Generally, ligand binding induces dimerization of RTK monomers, which induces auto-/transphosphorylation of tyrosine residues on the intracellular tails leading to the recruitment of adaptor proteins and modifying enzymes to promote and modulate various downstream signaling pathways. This chapter details easy, rapid, sensitive and versatile methods based on split Nanoluciferase complementation technology (NanoBiT) to monitor activation and modulation of two models of RTKs (EGFR and AXL) through the measurement of their dimerization and the recruitment of the adaptor protein Grb2 (SH2 domain-containing growth factor receptor-bound protein 2) and the receptor-modifying enzyme, the ubiquitin ligase Cbl.
摘要：
受体酪氨酸激酶（RTKs）是由多种生长因子激活的跨膜受体，细胞因子或激素。它们确保细胞过程中的多个角色，包括扩散，分化和生存。它们也是多种癌症类型发展和进展的关键驱动因素，并代表重要的药物靶标。一般来说,配体结合诱导RTK单体的二聚化，它诱导细胞内尾巴上酪氨酸残基的自动/转磷酸化，导致衔接蛋白和修饰酶的募集，以促进和调节各种下游信号通路。本章详细介绍了简单，快速,基于分裂纳米亮氨酸酶互补技术（NanoBiT）的灵敏且通用的方法，通过测量其二聚化和衔接蛋白Grb2（含SH2结构域的生长因子受体结合蛋白2）和受体修饰酶的募集来监测两种RTK（EGFR和AXL）模型的激活和调节，泛素连接酶Cbl.
