Cargando…

Ouabain-regulated phosphoproteome reveals molecular mechanisms for Na(+), K(+)–ATPase control of cell adhesion, proliferation, and survival

The ion pump Na(+), K(+)–ATPase (NKA) is a receptor for the cardiotonic steroid ouabain. Subsaturating concentration of ouabain triggers intracellular calcium oscillations, stimulates cell proliferation and adhesion, and protects from apoptosis. However, it is controversial whether ouabain-bound NKA...

Descripción completa

Detalles Bibliográficos
Autores principales: Panizza, Elena, Zhang, Liang, Fontana, Jacopo Maria, Hamada, Kozo, Svensson, Daniel, Akkuratov, Evgeny E., Scott, Lena, Mikoshiba, Katsuhiko, Brismar, Hjalmar, Lehtiö, Janne, Aperia, Anita
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Federation of American Societies for Experimental Biology 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6704450/
https://www.ncbi.nlm.nih.gov/pubmed/31199885
http://dx.doi.org/10.1096/fj.201900445R
_version_ 1783445508355784704
author Panizza, Elena
Zhang, Liang
Fontana, Jacopo Maria
Hamada, Kozo
Svensson, Daniel
Akkuratov, Evgeny E.
Scott, Lena
Mikoshiba, Katsuhiko
Brismar, Hjalmar
Lehtiö, Janne
Aperia, Anita
author_facet Panizza, Elena
Zhang, Liang
Fontana, Jacopo Maria
Hamada, Kozo
Svensson, Daniel
Akkuratov, Evgeny E.
Scott, Lena
Mikoshiba, Katsuhiko
Brismar, Hjalmar
Lehtiö, Janne
Aperia, Anita
author_sort Panizza, Elena
collection PubMed
description The ion pump Na(+), K(+)–ATPase (NKA) is a receptor for the cardiotonic steroid ouabain. Subsaturating concentration of ouabain triggers intracellular calcium oscillations, stimulates cell proliferation and adhesion, and protects from apoptosis. However, it is controversial whether ouabain-bound NKA is considered a signal transducer. To address this question, we performed a global analysis of protein phosphorylation in COS-7 cells, identifying 2580 regulated phosphorylation events on 1242 proteins upon 10- and 20-min treatment with ouabain. Regulated phosphorylated proteins include the inositol triphosphate receptor and stromal interaction molecule, which are essential for initiating calcium oscillations. Hierarchical clustering revealed that ouabain triggers a structured phosphorylation response that occurs in a well-defined, time-dependent manner and affects specific cellular processes, including cell proliferation and cell-cell junctions. We additionally identify regulation of the phosphorylation of several calcium and calmodulin–dependent protein kinases (CAMKs), including 2 sites of CAMK type II-γ (CAMK2G), a protein known to regulate apoptosis. To verify the significance of this result, CAMK2G was knocked down in primary kidney cells. CAMK2G knockdown impaired ouabain-dependent protection from apoptosis upon treatment with high glucose or serum deprivation. In conclusion, we establish NKA as the coordinator of a broad, tightly regulated phosphorylation response in cells and define CAMK2G as a downstream effector of NKA.—Panizza, E., Zhang, L., Fontana, J. M., Hamada, K., Svensson, D., Akkuratov, E. E., Scott, L., Mikoshiba, K., Brismar, H., Lehtiö, J., Aperia, A. Ouabain-regulated phosphoproteome reveals molecular mechanisms for Na(+), K(+)–ATPase control of cell adhesion, proliferation, and survival.
format Online
Article
Text
id pubmed-6704450
institution National Center for Biotechnology Information
language English
publishDate 2019
publisher Federation of American Societies for Experimental Biology
record_format MEDLINE/PubMed
spelling pubmed-67044502019-08-27 Ouabain-regulated phosphoproteome reveals molecular mechanisms for Na(+), K(+)–ATPase control of cell adhesion, proliferation, and survival Panizza, Elena Zhang, Liang Fontana, Jacopo Maria Hamada, Kozo Svensson, Daniel Akkuratov, Evgeny E. Scott, Lena Mikoshiba, Katsuhiko Brismar, Hjalmar Lehtiö, Janne Aperia, Anita FASEB J Research The ion pump Na(+), K(+)–ATPase (NKA) is a receptor for the cardiotonic steroid ouabain. Subsaturating concentration of ouabain triggers intracellular calcium oscillations, stimulates cell proliferation and adhesion, and protects from apoptosis. However, it is controversial whether ouabain-bound NKA is considered a signal transducer. To address this question, we performed a global analysis of protein phosphorylation in COS-7 cells, identifying 2580 regulated phosphorylation events on 1242 proteins upon 10- and 20-min treatment with ouabain. Regulated phosphorylated proteins include the inositol triphosphate receptor and stromal interaction molecule, which are essential for initiating calcium oscillations. Hierarchical clustering revealed that ouabain triggers a structured phosphorylation response that occurs in a well-defined, time-dependent manner and affects specific cellular processes, including cell proliferation and cell-cell junctions. We additionally identify regulation of the phosphorylation of several calcium and calmodulin–dependent protein kinases (CAMKs), including 2 sites of CAMK type II-γ (CAMK2G), a protein known to regulate apoptosis. To verify the significance of this result, CAMK2G was knocked down in primary kidney cells. CAMK2G knockdown impaired ouabain-dependent protection from apoptosis upon treatment with high glucose or serum deprivation. In conclusion, we establish NKA as the coordinator of a broad, tightly regulated phosphorylation response in cells and define CAMK2G as a downstream effector of NKA.—Panizza, E., Zhang, L., Fontana, J. M., Hamada, K., Svensson, D., Akkuratov, E. E., Scott, L., Mikoshiba, K., Brismar, H., Lehtiö, J., Aperia, A. Ouabain-regulated phosphoproteome reveals molecular mechanisms for Na(+), K(+)–ATPase control of cell adhesion, proliferation, and survival. Federation of American Societies for Experimental Biology 2019-09 2019-07-10 /pmc/articles/PMC6704450/ /pubmed/31199885 http://dx.doi.org/10.1096/fj.201900445R Text en © The Author(s) http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution 4.0 International (CC BY 4.0) (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Panizza, Elena
Zhang, Liang
Fontana, Jacopo Maria
Hamada, Kozo
Svensson, Daniel
Akkuratov, Evgeny E.
Scott, Lena
Mikoshiba, Katsuhiko
Brismar, Hjalmar
Lehtiö, Janne
Aperia, Anita
Ouabain-regulated phosphoproteome reveals molecular mechanisms for Na(+), K(+)–ATPase control of cell adhesion, proliferation, and survival
title Ouabain-regulated phosphoproteome reveals molecular mechanisms for Na(+), K(+)–ATPase control of cell adhesion, proliferation, and survival
title_full Ouabain-regulated phosphoproteome reveals molecular mechanisms for Na(+), K(+)–ATPase control of cell adhesion, proliferation, and survival
title_fullStr Ouabain-regulated phosphoproteome reveals molecular mechanisms for Na(+), K(+)–ATPase control of cell adhesion, proliferation, and survival
title_full_unstemmed Ouabain-regulated phosphoproteome reveals molecular mechanisms for Na(+), K(+)–ATPase control of cell adhesion, proliferation, and survival
title_short Ouabain-regulated phosphoproteome reveals molecular mechanisms for Na(+), K(+)–ATPase control of cell adhesion, proliferation, and survival
title_sort ouabain-regulated phosphoproteome reveals molecular mechanisms for na(+), k(+)–atpase control of cell adhesion, proliferation, and survival
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6704450/
https://www.ncbi.nlm.nih.gov/pubmed/31199885
http://dx.doi.org/10.1096/fj.201900445R
work_keys_str_mv AT panizzaelena ouabainregulatedphosphoproteomerevealsmolecularmechanismsfornakatpasecontrolofcelladhesionproliferationandsurvival
AT zhangliang ouabainregulatedphosphoproteomerevealsmolecularmechanismsfornakatpasecontrolofcelladhesionproliferationandsurvival
AT fontanajacopomaria ouabainregulatedphosphoproteomerevealsmolecularmechanismsfornakatpasecontrolofcelladhesionproliferationandsurvival
AT hamadakozo ouabainregulatedphosphoproteomerevealsmolecularmechanismsfornakatpasecontrolofcelladhesionproliferationandsurvival
AT svenssondaniel ouabainregulatedphosphoproteomerevealsmolecularmechanismsfornakatpasecontrolofcelladhesionproliferationandsurvival
AT akkuratovevgenye ouabainregulatedphosphoproteomerevealsmolecularmechanismsfornakatpasecontrolofcelladhesionproliferationandsurvival
AT scottlena ouabainregulatedphosphoproteomerevealsmolecularmechanismsfornakatpasecontrolofcelladhesionproliferationandsurvival
AT mikoshibakatsuhiko ouabainregulatedphosphoproteomerevealsmolecularmechanismsfornakatpasecontrolofcelladhesionproliferationandsurvival
AT brismarhjalmar ouabainregulatedphosphoproteomerevealsmolecularmechanismsfornakatpasecontrolofcelladhesionproliferationandsurvival
AT lehtiojanne ouabainregulatedphosphoproteomerevealsmolecularmechanismsfornakatpasecontrolofcelladhesionproliferationandsurvival
AT aperiaanita ouabainregulatedphosphoproteomerevealsmolecularmechanismsfornakatpasecontrolofcelladhesionproliferationandsurvival