PO.CH02.01 · 化学
相分离的RTK融合凝聚体通过广泛的蛋白招募和RTK串扰协调致癌信号
Phase-separated RTK fusion condensates orchestrate oncogenic signaling through extensive protein recruitment and RTK crosstalk
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
液-液相分离的RTK（受体酪氨酸激酶）融合凝聚体代表了一类新近鉴定的生物分子组装体，在驱动RTK融合蛋白的致癌功能中发挥关键作用。在本研究中，我们分离了NACC2-NTRK2（伏隔核相关蛋白2-神经营养受体酪氨酸激酶2）融合凝聚体，并应用LC-MS/MS结合IMAC（固定化金属亲和色谱）来分析其总蛋白质组和磷酸化蛋白质组。我们的分析揭示，该凝聚体通过激酶激活选择性招募了190种蛋白，影响诸如信号转导、转录调控、细胞分裂、细胞周期进程和代谢控制等关键细胞通路。荧光显微镜进一步表明，具有不同细胞功能的蛋白以不同方式与凝聚体相互作用。磷酸化蛋白质组学分析显示，激酶活性凝聚体使1,399种蛋白发生磷酸化，并促进RTK串扰，特别是与ephrin受体酪氨酸激酶的串扰，从而实现EPHA2（Ephrin受体A2）和EPHB4（Ephrin受体B4）的非配体依赖性激活。
查看英文原文 English abstract
Liquid-liquid phase-separated RTK (Receptor Tyrosine Kinase) fusion condensates represent a newly identified category of biomolecular assemblies that play a key role in driving the oncogenic functions of RTK fusion proteins. In this study, we isolated the NACC2-NTRK2 (Nucleus Accumbens-Associated Protein 2 - Neurotrophic Receptor Tyrosine Kinase 2) fusion condensate and applied LC-MS/MS along with IMAC (Immobilized Metal Affinity Chromatography) to profile its total proteome and phospho-proteome. Our analyses revealed that the condensate selectively recruits 190 proteins via kinase activation, impacting essential cellular pathways such as signal transduction, transcriptional regulation, cell division, cell-cycle progression, and metabolic control. Fluorescence microscopy further demonstrated that proteins with distinct cellular roles interact with the condensates in different ways. Phospho-proteomic profiling showed that the kinase active condensates bring phosphorylation to 1,399 proteins and facilitate RTK crosstalk, particularly with ephrin receptor tyrosine kinases, enabling ligand-independent activation of EPHA2 (Ephrin Receptor A2) and EPHB4 (Ephrin Receptor B4).
利益披露 Disclosure
W. Yang, None..
T. C. Whisenant, None..
D. J. Donoghue, None.