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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...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4477410/ https://www.ncbi.nlm.nih.gov/pubmed/26099953 http://dx.doi.org/10.1038/srep11133 |
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author | 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 |
author_facet | 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 |
author_sort | Röck, Ruth |
collection | PubMed |
description | 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. |
format | Online Article Text |
id | pubmed-4477410 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-44774102015-07-13 In-vivo detection of binary PKA network interactions upon activation of endogenous GPCRs 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 Sci Rep Article 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. Nature Publishing Group 2015-06-23 /pmc/articles/PMC4477410/ /pubmed/26099953 http://dx.doi.org/10.1038/srep11133 Text en Copyright © 2015, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article 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 In-vivo detection of binary PKA network interactions upon activation of endogenous GPCRs |
title | In-vivo detection of binary PKA network interactions upon activation of endogenous GPCRs |
title_full | In-vivo detection of binary PKA network interactions upon activation of endogenous GPCRs |
title_fullStr | In-vivo detection of binary PKA network interactions upon activation of endogenous GPCRs |
title_full_unstemmed | In-vivo detection of binary PKA network interactions upon activation of endogenous GPCRs |
title_short | In-vivo detection of binary PKA network interactions upon activation of endogenous GPCRs |
title_sort | in-vivo detection of binary pka network interactions upon activation of endogenous gpcrs |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4477410/ https://www.ncbi.nlm.nih.gov/pubmed/26099953 http://dx.doi.org/10.1038/srep11133 |
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