Using a kinase-inducible bimolecular switch to control enzyme activity in living cells

Curr Protoc Chem Biol. 2013;5(3):227-37. doi: 10.1002/9780470559277.ch130090.

Abstract

Molecular switches have been instrumental in the development of powerful and versatile genetic tools for directly probing biochemical processes, such as intracellular signaling, within their native contexts. A molecular switch can be broadly defined as a molecular system capable of existing in either of two states (e.g., conformations), which can be converted from one state to the other by a specific input stimulus. This protocol outlines a method for using a kinase-inducible bimolecular switch, along with live-cell fluorescence microscopy, to directly control and monitor the activity of a specific enzyme in living cells.

Keywords: biochemical perturbation; biosensor; fluorescence; live-cell imaging.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Line
  • Cell Physiological Phenomena / genetics*
  • DNA / genetics
  • Genes, Switch / genetics*
  • Humans
  • Indicators and Reagents
  • Microscopy, Fluorescence
  • Phosphotransferases / genetics*
  • Signal Transduction

Substances

  • Indicators and Reagents
  • DNA
  • Phosphotransferases