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The Use of FRET/FLIM to Study Proteins Interacting with Plant Receptor Kinases

  • Stefanie Weidtkamp-PetersEmail author
  • Yvonne StahlEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1621)

Abstract

The investigation of protein interactions in living plant tissue has become of increasing importance in recent years. A high spatial and temporal resolution for the observation of in vivo protein interaction is needed, e.g., in order to follow changes of plant receptor kinase interactions and complex formation over time. In vivo fluorescence or Förster resonance energy transfer (FRET) measurements allow for detailed analyses of interacting proteins in their natural environment at a subcellular level. Especially FRET-FLIM (fluorescence lifetime imaging microscopy) measurements provide an extremely powerful and reliable tool meeting the demands for investigating in vivo protein interaction quantitatively and with high precision. Here, we will describe in detail how to practically perform in vivo FRET measurements of receptor kinases in plants and discuss potential pitfalls and points of consideration.

Key words

FRET APB FLIM Fluorescent proteins Protein interaction 

Notes

Acknowledgments

We acknowledge the Deutsche Forschungsgesellschaft (DFG) for financial support to Y.S. by grant STA12/12 1-1 and S.W.P. by grant WE53/43 1-1. We are thankful to Steffen Köhler for taking photographs of N. benthamiana leaves and Rüdiger Simon for critical reading of the manuscript.

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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Center for Advanced Imaging (CAi)Heinrich-Heine UniversityDüsseldorfGermany
  2. 2.Institute for Developmental GeneticsHeinrich-Heine UniversityDüsseldorfGermany

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