Optogenetic control of protein binding using light-switchable nanobodies

Agnieszka A. Gil, César Carrasco-López, Liyuan Zhu, Evan M. Zhao, Pavithran T. Ravindran, Maxwell Z. Wilson, Alexander G. Goglia, José L. Avalos () and Jared E. Toettcher ()
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Agnieszka A. Gil: Princeton University
César Carrasco-López: Princeton University
Liyuan Zhu: Princeton University
Evan M. Zhao: Princeton University
Pavithran T. Ravindran: Princeton University
Maxwell Z. Wilson: Princeton University
Alexander G. Goglia: Princeton University
José L. Avalos: Princeton University
Jared E. Toettcher: Princeton University

Nature Communications, 2020, vol. 11, issue 1, 1-12

Abstract: Abstract A growing number of optogenetic tools have been developed to reversibly control binding between two engineered protein domains. In contrast, relatively few tools confer light-switchable binding to a generic target protein of interest. Such a capability would offer substantial advantages, enabling photoswitchable binding to endogenous target proteins in cells or light-based protein purification in vitro. Here, we report the development of opto-nanobodies (OptoNBs), a versatile class of chimeric photoswitchable proteins whose binding to proteins of interest can be enhanced or inhibited upon blue light illumination. We find that OptoNBs are suitable for a range of applications including reversibly binding to endogenous intracellular targets, modulating signaling pathway activity, and controlling binding to purified protein targets in vitro. This work represents a step towards programmable photoswitchable regulation of a wide variety of target proteins.

Date: 2020
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DOI: 10.1038/s41467-020-17836-8

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