cAMP Signaling pp 151-166 | Cite as

Screening for Small Molecule Disruptors of AKAP–PKA Interactions

  • Carolin Schächterle
  • Frank Christian
  • João Miguel Parente Fernandes
  • Enno KlussmannEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1294)


Protein–protein interactions (PPIs) are highly specific and diverse. Their selective inhibition with peptides, peptidomimetics, or small molecules allows determination of functions of individual PPIs. Moreover, inhibition of disease-associated PPIs may lead to new concepts for the treatment of diseases with an unmet medical need. Protein kinase A (PKA) is an ubiquitously expressed protein kinase that controls a plethora of cellular functions. A-kinase anchoring proteins (AKAPs) are multivalent scaffolding proteins that directly interact with PKA. AKAPs spatially and temporally restrict PKA activity to defined cellular compartments and thereby contribute to the specificity of PKA signaling. However, it is largely unknown which of the plethora of PKA-dependent signaling events involve interactions of PKA with AKAPs. Moreover, AKAP–PKA interactions appear to play a role in a variety of cardiovascular, neuronal, and inflammatory diseases, but it is unclear whether these interactions are suitable drug targets. Here we describe an enzyme-linked immunosorbent assay (ELISA) for the screening of small molecule libraries for inhibitors of AKAP–PKA interactions. In addition, we describe a homogenous time-resolved fluorescence (HTRF) assay for use in secondary validation screens. Small molecule inhibitors are invaluable molecular tools for elucidating the functions of AKAP–PKA interactions and may eventually lead to new concepts for the treatment of diseases where AKAP–PKA interactions represent potential drug targets.


Protein kinase A (PKA) A-kinase anchoring protein (AKAP) Inhibitory peptides Nonpeptidic helix mimetics Small molecules Homogenous time-resolved fluorescence (HTRF) assay Enzyme-linked immunosorbent assay (ELISA) 



We thank Sylvia Niquet for technical assistance. This work was supported by the Deutsche Forschungsgemeinschaft (DFG KL1415/4-2), the Else Kröner-Fresenius-Stiftung (2013_A145), and the German-Israeli Foundation (I-1210-286.13/2012).


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Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Carolin Schächterle
    • 1
  • Frank Christian
    • 2
  • João Miguel Parente Fernandes
    • 1
  • Enno Klussmann
    • 1
    Email author
  1. 1.Max Delbruck Center for Molecular Medicine (MDC) Berlin-BuchBerlinGermany
  2. 2.Institute of Cardiovascular and Medical SciencesUniversity of GlasgowGlasgowUK

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