BiFC Assay to Detect Calmodulin Binding to Plant Receptor Kinases

  • Cornelia Fischer
  • Margret Sauter
  • Petra Dietrich
Part of the Methods in Molecular Biology book series (MIMB, volume 1621)


Plant receptor-like kinases (RLKs) are regulated at various levels including posttranscriptional modification and interaction with regulatory proteins. Calmodulin (CaM) is a calcium-sensing protein that was shown to bind to some RLKs such as the PHYTOSULFOKINE RECEPTOR1 (PSKR1). The CaM-binding site is embedded in subdomain VIa of the kinase domain. It is possible that many more of RLKs interact with CaM than previously described. To unequivocally confirm CaM binding, several methods exist. Bimolecular fluorescence complementation (BiFC) and pull-down assays have been successfully used to study CaM binding to PSKR1 and are described in this chapter (BiFC) and in  Chapter 15 (pull down). The two methods are complementary. BiFC is useful to show localization and interaction of soluble as well as of membrane-bound proteins in planta.

Key words

Agrobacterium-mediated transformation of N. benthamiana BiFC Confocal laser scanning microscope Calmodulin Peptide receptor kinase Protein binding 



We would like to thank Heiner Busch for photography and the Deutsche Forschungsgemeinschaft for financial support.


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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Cornelia Fischer
    • 1
  • Margret Sauter
    • 2
  • Petra Dietrich
    • 1
  1. 1.Molecular Plant Physiology, Department of BiologyUniversity of Erlangen-NurembergErlangenGermany
  2. 2.Plant Developmental Biology and Plant PhysiologyUniversity of KielKielGermany

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