@article{e839ba6648914792b56dcfff88d78d07,
title = "Bacterial Phytochrome as a Scaffold for Engineering of Receptor Tyrosine Kinases Controlled with Near-Infrared Light",
abstract = "Optically controlled receptor tyrosine kinases (opto-RTKs) allow regulation of RTK signaling using light. Until recently, the majority of opto-RTKs were activated with blue-green light. Fusing a photosensory core module of Deinococcus radiodurans bacterial phytochrome (DrBphP-PCM) to the kinase domains of neurotrophin receptors resulted in opto-RTKs controlled with light above 650 nm. To expand this engineering approach to RTKs of other families, here we combined the DrBpP-PCM with the cytoplasmic domains of EGFR and FGFR1. The resultant Dr-EGFR and Dr-FGFR1 opto-RTKs are rapidly activated with near-infrared and inactivated with far-red light. The opto-RTKs efficiently trigger ERK1/2, PI3K/Akt, and PLCγ signaling. Absence of spectral crosstalk between the opto-RTKs and green fluorescent protein-based biosensors enables simultaneous Dr-FGFR1 activation and detection of calcium transients. Action mechanism of the DrBphP-PCM-based opto-RTKs is considered using the available RTK structures. DrBphP-PCM represents a versatile scaffold for engineering of opto-RTKs that are reversibly regulated with far-red and near-infrared light.",
keywords = "DrBphP, EGFR, FGFR1, bacteriophytochrome, opto-RTK",
author = "Leopold, {Anna V.} and Sergei Pletnev and Verkhusha, {Vladislav V.}",
note = "Funding Information: We thank J. Ihalainen (University of Jyv{\"a}skyl{\"a}, Finland) for the DrBphP gene and D. Lindholm (University of Helsinki, Finland) for the cell lines. This work was supported by grants GM122567 and NS103573 from the National Institutes of Health (NIH) and 322226 from the Academy of Finland . This project was also funded in part with US federal funds from the Frederick National Laboratory for Cancer Research , NIH contract HHSN261200800001E, the Intramural Research Program of the NIH, Frederick National Laboratory, Center for Cancer Research . Funding Information: We thank J. Ihalainen (University of Jyv?skyl?, Finland) for the DrBphP gene and D. Lindholm (University of Helsinki, Finland) for the cell lines. This work was supported by grants GM122567 and NS103573 from the National Institutes of Health (NIH) and 322226 from the Academy of Finland. This project was also funded in part with US federal funds from the Frederick National Laboratory for Cancer Research, NIH contract HHSN261200800001E, the Intramural Research Program of the NIH, Frederick National Laboratory, Center for Cancer Research. A.V.L. engineered and characterized the opto-RTKs. S.P. performed the structural modeling. V.V.V. planned and directed the project and together with A.V.L. and S.P. designed the experiments, analyzed the data, and wrote the manuscript. None. Publisher Copyright: {\textcopyright} 2020 Elsevier Ltd",
year = "2020",
month = jun,
day = "12",
doi = "10.1016/j.jmb.2020.04.005",
language = "English (US)",
volume = "432",
pages = "3749--3760",
journal = "Journal of Molecular Biology",
issn = "0022-2836",
publisher = "Academic Press Inc.",
number = "13",
}