Neurotrophin receptor tyrosine kinases regulated with near-infrared light

• Published in: Nature communications 2019-03, Vol.10 (1), p.1129-1129, Article 1129
• Main Authors: Leopold, Anna V., Chernov, Konstantin G., Shemetov, Anton A., Verkhusha, Vladislav V.
• Format: Article
• Language: English
• Subjects: 13; 14; 631/1647/2253; 631/1647/338/469; 631/92/275; Activation; Animals; Apoptosis; Apoptosis - genetics; Apoptosis - radiation effects; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; Calcium; Catalysis; Cell Line, Tumor; Crosstalk; Domains; Epithelial Cells - cytology; Epithelial Cells - metabolism; Epithelial Cells - radiation effects; Gene Expression Regulation; Glioblastoma; Growth factors; HeLa Cells; Humanities and Social Sciences; Humans; I.R. radiation; Infrared radiation; Infrared Rays; Intracellular signalling; Kinases; Ligands; Light Signal Transduction; Mice; multidisciplinary; Multiplexing; Nerve Growth Factors - genetics; Nerve Growth Factors - metabolism; Neuroblastoma; Neuroglia - cytology; Neuroglia - metabolism; Neuroglia - radiation effects; Neurons - cytology; Neurons - metabolism; Neurons - radiation effects; Neurotrophic factors; Optical control; Optogenetics - methods; Phytochrome - genetics; Phytochrome - metabolism; Protein Engineering; Proteins; Receptor, trkA - genetics; Receptor, trkA - metabolism; Receptor, trkB - genetics; Receptor, trkB - metabolism; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; Science; Science (multidisciplinary); Stability; Translocation; TrkA protein; TrkA receptors; TrkB receptors; Tropomyosin; Tyrosine
• ISSN: 2041-1723; 2041-1723
• 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. Optical control over receptor tyrosine kinase (RTK) activity can find application in both basic biology and synthetic biology. Here the authors combine the photosensory module of DrBphP bacterial phytochrome with the Trk family of RTKs to obtain Trks that are reversibly switchable with near-infrared and far-red light.