In-vivo detection of binary PKA network interactions upon activation of endogenous GPCRs

Membrane receptor-sensed input signals affect and modulate intracellular protein-protein interactions (PPIs). Consequent changes occur to the compositions of protein complexes, protein localization and intermolecular binding affinities. Alterations of compartmentalized PPIs emanating from certain de...

Full description

Saved in:
Bibliographic Details
Published in:Scientific reports 2015-06, Vol.5 (1), p.11133-11133, Article 11133
Main Authors: Röck, Ruth, Bachmann, Verena, Bhang, Hyo-Eun C, Malleshaiah, Mohan, Raffeiner, Philipp, Mayrhofer, Johanna E, Tschaikner, Philipp M, Bister, Klaus, Aanstad, Pia, Pomper, Martin G, Michnick, Stephen W, Stefan, Eduard
Format: Article
Language:English
Subjects:
Animals
Biosensing Techniques
Cancer
Cell Line, Tumor
Complementation
Cyclic AMP-Dependent Protein Kinases - metabolism
Embryo, Nonmammalian - metabolism
Embryos
G protein-coupled receptors
Genes, Reporter
HEK293 Cells
Humans
Kinases
Localization
Mice
Osteosarcoma - metabolism
Protein Binding
Protein interaction
Protein Interaction Mapping
Protein kinase A
Proteins
Receptors, G-Protein-Coupled - metabolism
Saccharomyces cerevisiae - metabolism
Xenograft Model Antitumor Assays
Xenografts
Zebrafish - embryology
Zebrafish - metabolism
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Membrane receptor-sensed input signals affect and modulate intracellular protein-protein interactions (PPIs). Consequent changes occur to the compositions of protein complexes, protein localization and intermolecular binding affinities. Alterations of compartmentalized PPIs emanating from certain deregulated kinases are implicated in the manifestation of diseases such as cancer. Here we describe the application of a genetically encoded Protein-fragment Complementation Assay (PCA) based on the Renilla Luciferase (Rluc) enzyme to compare binary PPIs of the spatially and temporally controlled protein kinase A (PKA) network in diverse eukaryotic model systems. The simplicity and sensitivity of this cell-based reporter allows for real-time recordings of mutually exclusive PPIs of PKA upon activation of selected endogenous G protein-coupled receptors (GPCRs) in cancer cells, xenografts of mice, budding yeast, and zebrafish embryos. This extends the application spectrum of Rluc PCA for the quantification of PPI-based receptor-effector relationships in physiological and pathological model systems.
ISSN:2045-2322
2045-2322
DOI:10.1038/srep11133