TY - JOUR
T1 - Optogenetic control of protein binding using light-switchable nanobodies
AU - Gil, Agnieszka A.
AU - Carrasco-López, César
AU - Zhu, Liyuan
AU - Zhao, Evan M.
AU - Ravindran, Pavithran T.
AU - Wilson, Maxwell Z.
AU - Goglia, Alexander G.
AU - Avalos, José L.
AU - Toettcher, Jared E.
N1 - Funding Information:
We thank all members of the Toettcher and Avalos labs for helpful comments, and Nicole Neville for her initial opto-nanobody studies. We also thank the Biophysics Core Facility and Venu Vandavasi for help with the bio-layer interferometry measurements. This work was supported by NIH grant DP2EB024247 (to J.E.T.), the Pew Charitable Trusts, the U.S. DOE Office of Biological and Environmental Research, Genomic Science Program Award DESC0019363, NSF CAREER Award CBET-1751840, and a Camille Dreyfus Teacher-Scholar Award (to J.L.A.), NIH Fellowship F32GM128304 (to A.A.G.), and NIH Fellowship F30CA206408 (to A.G.G.).
PY - 2020/12/1
Y1 - 2020/12/1
N2 - 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.
AB - 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.
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U2 - 10.1038/s41467-020-17836-8
DO - 10.1038/s41467-020-17836-8
M3 - Article
C2 - 32792536
AN - SCOPUS:85089411213
VL - 11
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
IS - 1
M1 - 4044
ER -