Wurz Anna I, Zheng Kevin S, Hughes Robert M
Department of Chemistry, East Carolina University, Greenville, North Carolina, United States.
Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina, United States.
bioRxiv. 2024 Feb 7:2024.02.06.579139. doi: 10.1101/2024.02.06.579139.
Eph receptors are ubiquitous class of transmembrane receptors that mediate cell-cell communication, proliferation, differentiation, and migration. EphA1 receptors specifically play an important role in angiogenesis, fetal development, and cancer progression; however, studies of this receptor can be challenging as its ligand, ephrinA1, binds and activates several EphA receptors simultaneously. Optogenetic strategies could be applied to circumvent this requirement for ligand activation and enable selective activation of the EphA1 subtype. In this work, we designed and tested several iterations of an optogenetic EphA1 - Cryptochrome 2 (Cry2) fusion, investigating their capacity to mimic EphA1-dependent signaling in response to light activation. We then characterized the key cell signaling target of MAPK phosphorylation activated in response to light stimulation. The optogenetic regulation of Eph receptor RTK signaling without the need for external stimulus promises to be an effective means of controlling individual Eph receptor-mediated activities and creates a path forward for the identification of new Eph-dependent functions.
Eph受体是一类普遍存在的跨膜受体，介导细胞间通讯、增殖、分化和迁移。EphA1受体在血管生成、胎儿发育和癌症进展中特别发挥重要作用；然而，对该受体的研究可能具有挑战性，因为其配体ephrinA1会同时结合并激活多种EphA受体。光遗传学策略可用于规避这种对配体激活的需求，并实现对EphA1亚型的选择性激活。在这项工作中，我们设计并测试了几种光遗传学EphA1 - 隐花色素2（Cry2）融合体的迭代版本，研究它们在光激活时模拟EphA1依赖性信号传导的能力。然后，我们对响应光刺激而激活的MAPK磷酸化的关键细胞信号靶点进行了表征。无需外部刺激即可对Eph受体RTK信号进行光遗传学调控，有望成为控制单个Eph受体介导的活动的有效手段，并为鉴定新的Eph依赖性功能开辟一条道路。
