Sensitizing plant protein kinases to specific inhibition by atp-competitive molecules

Dor Salomon, Chao Zhang, Kevan M. Shokat, Guido Sessa

Research output: Chapter in Book/Report/Conference proceedingChapterpeer-review

Abstract

The highly conserved nature of the protein kinase catalytic domain and the low permeability of plant cell membranes pose a challenge to the development of specific inhibitors that target individual protein kinases in vivo. Here, we describe a chemical-genetic approach to specifically sensitize individual plant kinases to cell-permeable small molecules that do not inhibit wild-type kinases. In this approach, a single amino-acid substitution is introduced in the ATP-binding site of the enzyme enabling specific binding of ATP-competitive molecules. Cell-permeable molecules can then be used to specifically target the sensitized allele in transgenic Arabidopsis thaliana plants that do not express the wild-type form of the kinase. This strategy provides a useful tool for the functional characterization of protein kinases in planta and for the dissection of the signaling pathways in which they are involved.

Original languageEnglish
Title of host publicationPlant Kinases
Subtitle of host publicationMethods and Protocols
PublisherHumana Press Inc.
Pages185-197
Number of pages13
ISBN (Print)9781617792632
DOIs
StatePublished - 2011

Publication series

NameMethods in Molecular Biology
Volume779
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • ATP-competitive molecules
  • Analog-sensitive
  • Arabidopsis
  • Gatekeeper
  • Kinase activity assay
  • PP1
  • Protein kinase
  • Transgenic plants

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