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Impact of kinase activating and inactivating patient mutations on binary PKA interactions
The second messenger molecule cAMP links extracellular signals to intracellular responses. The main cellular cAMP effector is the compartmentalized protein kinase A (PKA). Upon receptor initiated cAMP-mobilization, PKA regulatory subunits (R) bind cAMP thereby triggering dissociation and activation...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2015
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4539479/ https://www.ncbi.nlm.nih.gov/pubmed/26347651 http://dx.doi.org/10.3389/fphar.2015.00170 |
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author | Röck, Ruth Mayrhofer, Johanna E. Bachmann, Verena Stefan, Eduard |
author_facet | Röck, Ruth Mayrhofer, Johanna E. Bachmann, Verena Stefan, Eduard |
author_sort | Röck, Ruth |
collection | PubMed |
description | The second messenger molecule cAMP links extracellular signals to intracellular responses. The main cellular cAMP effector is the compartmentalized protein kinase A (PKA). Upon receptor initiated cAMP-mobilization, PKA regulatory subunits (R) bind cAMP thereby triggering dissociation and activation of bound PKA catalytic subunits (PKAc). Mutations in PKAc or RIa subunits manipulate PKA dynamics and activities which contribute to specific disease patterns. Mutations activating cAMP/PKA signaling contribute to carcinogenesis or hormone excess, while inactivating mutations cause hormone deficiency or resistance. Here we extended the application spectrum of a Protein-fragment Complementation Assay based on the Renilla Luciferase to determine binary protein:protein interactions (PPIs) of the PKA network. We compared time- and dose-dependent influences of cAMP-elevation on mutually exclusive PPIs of PKAc with the phosphotransferase inhibiting RIIb and RIa subunits and the protein kinase inhibitor peptide (PKI). We analyzed PKA dynamics following integration of patient mutations into PKAc and RIa. We observed that oncogenic modifications of PKAc(L206R) and RIa(Δ184-236) as well as rare disease mutations in RIa(R368X) affect complex formation of PKA and its responsiveness to cAMP elevation. With the cell-based PKA PPI reporter platform we precisely quantified the mechanistic details how inhibitory PKA interactions and defined patient mutations contribute to PKA functions. |
format | Online Article Text |
id | pubmed-4539479 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2015 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-45394792015-09-07 Impact of kinase activating and inactivating patient mutations on binary PKA interactions Röck, Ruth Mayrhofer, Johanna E. Bachmann, Verena Stefan, Eduard Front Pharmacol Pharmacology The second messenger molecule cAMP links extracellular signals to intracellular responses. The main cellular cAMP effector is the compartmentalized protein kinase A (PKA). Upon receptor initiated cAMP-mobilization, PKA regulatory subunits (R) bind cAMP thereby triggering dissociation and activation of bound PKA catalytic subunits (PKAc). Mutations in PKAc or RIa subunits manipulate PKA dynamics and activities which contribute to specific disease patterns. Mutations activating cAMP/PKA signaling contribute to carcinogenesis or hormone excess, while inactivating mutations cause hormone deficiency or resistance. Here we extended the application spectrum of a Protein-fragment Complementation Assay based on the Renilla Luciferase to determine binary protein:protein interactions (PPIs) of the PKA network. We compared time- and dose-dependent influences of cAMP-elevation on mutually exclusive PPIs of PKAc with the phosphotransferase inhibiting RIIb and RIa subunits and the protein kinase inhibitor peptide (PKI). We analyzed PKA dynamics following integration of patient mutations into PKAc and RIa. We observed that oncogenic modifications of PKAc(L206R) and RIa(Δ184-236) as well as rare disease mutations in RIa(R368X) affect complex formation of PKA and its responsiveness to cAMP elevation. With the cell-based PKA PPI reporter platform we precisely quantified the mechanistic details how inhibitory PKA interactions and defined patient mutations contribute to PKA functions. Frontiers Media S.A. 2015-08-18 /pmc/articles/PMC4539479/ /pubmed/26347651 http://dx.doi.org/10.3389/fphar.2015.00170 Text en Copyright © 2015 Röck, Mayrhofer, Bachmann and Stefan. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Pharmacology Röck, Ruth Mayrhofer, Johanna E. Bachmann, Verena Stefan, Eduard Impact of kinase activating and inactivating patient mutations on binary PKA interactions |
title | Impact of kinase activating and inactivating patient mutations on binary PKA interactions |
title_full | Impact of kinase activating and inactivating patient mutations on binary PKA interactions |
title_fullStr | Impact of kinase activating and inactivating patient mutations on binary PKA interactions |
title_full_unstemmed | Impact of kinase activating and inactivating patient mutations on binary PKA interactions |
title_short | Impact of kinase activating and inactivating patient mutations on binary PKA interactions |
title_sort | impact of kinase activating and inactivating patient mutations on binary pka interactions |
topic | Pharmacology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4539479/ https://www.ncbi.nlm.nih.gov/pubmed/26347651 http://dx.doi.org/10.3389/fphar.2015.00170 |
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