title: Screening for small molecule disruptors of AKAP-PKA interactions.
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.
authors.affiliation: Max Delbruck Center for Molecular Medicine (MDC) Berlin-Buch, Robert-Rössle-Str. 10, Berlin, 13125, Germany.
authors.forename: Carolin
authors.initials: C
authors.lastname: Schächterle
authors.forename: Frank
authors.initials: F
authors.lastname: Christian
authors.forename: João Miguel Parente
authors.initials: JM
authors.lastname: Fernandes
authors.forename: Enno
authors.initials: E
authors.lastname: Klussmann
mesh_terms: D054758:A Kinase Anchor Proteins / Q000737:chemistry / Q000378:metabolism*; D017868:Cyclic AMP-Dependent Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005453:Fluorescence; D011485:Protein Binding / Q000187:drug effects; D015398:Signal Transduction; D054852:Small Molecule Libraries / Q000737:chemistry* / Q000494:pharmacology
chemical_list: D054758:A Kinase Anchor Proteins; D054852:Small Molecule Libraries; D017868:Cyclic AMP-Dependent Protein Kinases