TY - JOUR
T1 - Neurotrophin receptor tyrosine kinases regulated with near-infrared light
AU - Leopold, Anna V.
AU - Chernov, Konstantin G.
AU - Shemetov, Anton A.
AU - Verkhusha, Vladislav V.
N1 - Funding Information:
We thank J. Ihalainen (University of Jyväskylä, Finland) for the DrBphP gene, D. Lindholm (University of Helsinki, Finland) for the cell lines, S. Pletnev (Frederick National Laboratory for Cancer Research, USA) for the proposed mechanism of Dr-Trk operation, and Y. Sidorova, J. Umemori, O. Oliinyk, E. Omelina, and T. Redchuk (all from University of Helsinki, Finland) for the useful discussions. We also thank the staff of Biomedicum Imaging Unit, Biomedicum Flow Cytometry Facility, Neuronal Cell Culture Unit, AAV Gene Transfer Facility (all of Medicum), and High-Throughput Screening Unit (of Finland Institute for Molecular Medicine) of University of Helsinki, Finland, for the technical assistance. This work was supported by grants GM122567 and NS103573 from the US National Institutes of Health (NIH) and ERC-2013-ADG-340233 from the EU FP7 program.
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - 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.
AB - 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.
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U2 - 10.1038/s41467-019-08988-3
DO - 10.1038/s41467-019-08988-3
M3 - Article
C2 - 30850602
AN - SCOPUS:85062617068
SN - 2041-1723
VL - 10
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 1129
ER -