Tools for controlling protein interactions using light

Curr Protoc Cell Biol. 2014 Sep 2:64:17.16.1-20. doi: 10.1002/0471143030.cb1716s64.

Abstract

Genetically encoded actuators that allow control of protein-protein interactions using light, termed 'optical dimerizers', are emerging as new tools for experimental biology. In recent years, numerous new and versatile dimerizer systems have been developed. Here we discuss the design of optical dimerizer experiments, including choice of a dimerizer system, photoexcitation sources, and the coordinate use of imaging reporters. We provide detailed protocols for experiments using two dimerization systems we previously developed, CRY2/CIB and UVR8/UVR8, for use in controlling transcription, protein localization, and protein secretion using light. Additionally, we provide instructions and software for constructing a pulse-controlled LED device for use in experiments requiring extended light treatments.

Keywords: UVR8; cryptochrome; optogenetics; photoreceptor; protein interactions; protein secretion.

Publication types

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

MeSH terms

  • Animals
  • Arabidopsis Proteins / chemistry*
  • Arabidopsis Proteins / genetics
  • COS Cells
  • Chlorocebus aethiops
  • Chromosomal Proteins, Non-Histone / chemistry*
  • Chromosomal Proteins, Non-Histone / genetics
  • Cryptochromes / chemistry*
  • Cryptochromes / genetics
  • HeLa Cells
  • Humans
  • Light*
  • Protein Multimerization*

Substances

  • Arabidopsis Proteins
  • CRY2 protein, Arabidopsis
  • Chromosomal Proteins, Non-Histone
  • Cryptochromes
  • Uvr8 protein, Arabidopsis