Medicum, Faculty of Medicine, University of Helsinki, Helsinki, 00290, Finland.
Department of Anatomy and Structural Biology, and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY, 10461, USA.
Nat Commun. 2019 Mar 8;10(1):1129. doi: 10.1038/s41467-019-08988-3.
Optical control over the activity of receptor tyrosine kinases (RTKs) provides an efficient way to reversibly and non-invasively map their functions. We combined catalytic domains of Trk (tropomyosin receptor kinase) family of RTKs, naturally activated by neurotrophins, with photosensory core module of DrBphP bacterial phytochrome to develop opto-kinases, termed Dr-TrkA and Dr-TrkB, reversibly switchable on and off with near-infrared and far-red light. We validated Dr-Trk ability to reversibly light-control several RTK pathways, calcium level, and demonstrated that their activation triggers canonical Trk signaling. Dr-TrkA induced apoptosis in neuroblastoma and glioblastoma, but not in other cell types. Absence of spectral crosstalk between Dr-Trks and blue-light-activatable LOV-domain-based translocation system enabled intracellular targeting of Dr-TrkA independently of its activation, additionally modulating Trk signaling. Dr-Trks have several superior characteristics that make them the opto-kinases of choice for regulation of RTK signaling: high activation range, fast and reversible photoswitching, and multiplexing with visible-light-controllable optogenetic tools.
光学控制受体酪氨酸激酶 (RTKs) 的活性提供了一种高效的方法，可以可逆且非侵入性地映射它们的功能。我们将神经营养因子自然激活的 Trk（原肌球蛋白受体激酶）家族 RTKs 的催化结构域与细菌叶绿素光敏核心模块 DrBphP 结合，开发了光激酶，称为 Dr-TrkA 和 Dr-TrkB，可通过近红外和远红光进行可逆开关控制。我们验证了 Dr-Trk 可逆光控几种 RTK 途径、钙水平的能力，并证明其激活触发了典型的 Trk 信号转导。Dr-TrkA 诱导神经母细胞瘤和神经胶质瘤凋亡，但不诱导其他细胞类型凋亡。Dr-Trks 与蓝光激活的 LOV 结构域基于转位系统之间不存在光谱串扰，使得 Dr-TrkA 能够独立于其激活进行细胞内靶向，并且还可以调节 Trk 信号转导。Dr-Trks 具有几个优越的特性，使其成为调控 RTK 信号的光激酶的首选：高激活范围、快速且可逆的光开关以及与可见光可控光遗传学工具的多路复用。
