Design and Application of Light-Regulated Receptor Tyrosine Kinases

Stephanie Kainrath, Harald Janovjak

Research output: Chapter in Book/Report/Conference proceedingChapter (Book)Otherpeer-review

Abstract

Understanding how the activity of membrane receptors and cellular signaling pathways shapes cell behavior is of fundamental interest in basic and applied research. Reengineering receptors to react to light instead of their cognate ligands allows for generating defined signaling inputs with high spatial and temporal precision and facilitates the dissection of complex signaling networks. Here, we describe fundamental considerations in the design of light-regulated receptor tyrosine kinases (Opto-RTKs) and appropriate control experiments. We also introduce methods for transient receptor expression in HEK293 cells, quantitative assessment of signaling activity in reporter gene assays, semiquantitative assessment of (in)activation time courses through Western blot (WB) analysis, and easy to implement light stimulation hardware.

Original languageEnglish
Title of host publicationPhotoswitching Proteins
Subtitle of host publicationMethods and Protocols
EditorsDominik Niopek
Place of PublicationNew York, NY
PublisherHumana Press
Chapter16
Pages233-246
Number of pages14
ISBN (Electronic)9781071607558
ISBN (Print)9781071607541
DOIs
Publication statusPublished - 2020

Publication series

NameMethods in Molecular Biology
Volume2173
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Luciferase assay
  • Optogenetics
  • Photoreceptor
  • Receptor tyrosine kinase
  • Reporter gene assay
  • Transient transfection
  • Western blot

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