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Screening for small molecule disruptors of AKAP-PKA interactions

Item Type:Article
Title:Screening for small molecule disruptors of AKAP-PKA interactions
Creators Name:Schächterle, C. and Christian, F. and Fernandes, J.M.P. and Klussmann, E.
Abstract: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.
Keywords: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)
Source:Methods in Molecular Biology
Series Name:Methods in Molecular Biology
Title of Book:Screening for Small Molecule Disruptors of AKAP–PKA Interactions : Methods and Protocols
Publisher:Springer / Humana Press
Page Range:151-166
Date:2 March 2015
Official Publication:https://doi.org/10.1007/978-1-4939-2537-7_12
PubMed:View item in PubMed

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