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Disruptors of AKAP-dependent protein-protein interactions

Item Type:Article
Title:Disruptors of AKAP-dependent protein-protein interactions
Creators Name:Walker-Gray, R. and Pallien, T. and Miller, D.C. and Oder, A. and Neuenschwander, M. and von Kries, J.P. and Diecke, S. and Klussmann, E.
Abstract:A-kinase anchoring proteins (AKAPs) are a family of multivalent scaffolding proteins. They engage in direct protein-protein interactions with protein kinases, kinase substrates and further signaling molecules. Each AKAP interacts with a specific set of protein interaction partners and such sets can vary between different cellular compartments and cells. Thus, AKAPs can coordinate signal transduction processes spatially and temporally in defined cellular environments. AKAP-dependent protein-protein interactions are involved in a plethora of physiological processes, including processes in the cardiovascular, nervous, and immune system. Dysregulation of AKAPs and their interactions is associated with or causes widespread diseases, for example, cardiac diseases such as heart failure. However, there are profound shortcomings in understanding functions of specific AKAP-dependent protein-protein interactions. In part, this is due to the lack of agents for specifically targeting defined protein-protein interactions. Peptidic and non-peptidic inhibitors are invaluable molecular tools for elucidating the functions of AKAP-dependent protein-protein interactions. In addition, such interaction disruptors may pave the way to new concepts for the treatment of diseases where AKAP-dependent protein-protein interactions constitute potential drug targets.Here we describe screening approaches for the identification of small molecule disruptors of AKAP-dependent protein-protein interactions. Examples include interactions of AKAP18 and protein kinase A (PKA) and of AKAP-Lbc and RhoA. We discuss a homogenous time-resolved fluorescence (HTRF) and an AlphaScreen® assay for small molecule library screening and human induced pluripotent stem cell-derived cardiac myocytes (hiPSC-CMs) as a cell system for the characterization of identified hits.
Keywords:Protein Kinase A (PKA), A-kinase Anchoring Protein (AKAP), AKAP18, AKAP-Lbc, Inhibitory Peptides, Non-Peptidic Helix Mimetics, Small Molecules, Homogenous Time-Resolved Fluorescence (HTRF) Assay, AlphaScreen®, Calcium Imaging, Line Scan Imaging, Human Induced Pluripotent Stem Cells (hiPSCs)
Source:Methods in Molecular Biology
Series Name:Methods in Molecular Biology
Title of Book:cAMP signaling : methods and protocols
Publisher:Springer / Humana Press
Page Range:117-139
Official Publication:https://doi.org/10.1007/978-1-0716-2245-2_8
PubMed:View item in PubMed

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