Optogenetic control of protein binding using light-switchable nanobodies

Nat Commun. 2020 Aug 13;11(1):4044. doi: 10.1038/s41467-020-17836-8.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Animals
  • HEK293 Cells
  • Humans
  • Mice
  • NIH 3T3 Cells
  • Optogenetics / methods*
  • Protein Binding
  • Protein Transport / physiology
  • Signal Transduction / physiology
  • Synthetic Biology / methods*