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The human Na(+)/H(+) exchanger 1 is a membrane scaffold protein for extracellular signal-regulated kinase 2

BACKGROUND: Extracellular signal-regulated kinase 2 (ERK2) is an S/T kinase with more than 200 known substrates, and with critical roles in regulation of cell growth and differentiation and currently no membrane proteins have been linked to ERK2 scaffolding. METHODS AND RESULTS: Here, we identify th...

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Autores principales: Hendus-Altenburger, Ruth, Pedraz-Cuesta, Elena, Olesen, Christina W., Papaleo, Elena, Schnell, Jeff A., Hopper, Jonathan T. S., Robinson, Carol V., Pedersen, Stine F., Kragelund, Birthe B.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4833948/
https://www.ncbi.nlm.nih.gov/pubmed/27083547
http://dx.doi.org/10.1186/s12915-016-0252-7
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author Hendus-Altenburger, Ruth
Pedraz-Cuesta, Elena
Olesen, Christina W.
Papaleo, Elena
Schnell, Jeff A.
Hopper, Jonathan T. S.
Robinson, Carol V.
Pedersen, Stine F.
Kragelund, Birthe B.
author_facet Hendus-Altenburger, Ruth
Pedraz-Cuesta, Elena
Olesen, Christina W.
Papaleo, Elena
Schnell, Jeff A.
Hopper, Jonathan T. S.
Robinson, Carol V.
Pedersen, Stine F.
Kragelund, Birthe B.
author_sort Hendus-Altenburger, Ruth
collection PubMed
description BACKGROUND: Extracellular signal-regulated kinase 2 (ERK2) is an S/T kinase with more than 200 known substrates, and with critical roles in regulation of cell growth and differentiation and currently no membrane proteins have been linked to ERK2 scaffolding. METHODS AND RESULTS: Here, we identify the human Na(+)/H(+) exchanger 1 (hNHE1) as a membrane scaffold protein for ERK2 and show direct hNHE1-ERK1/2 interaction in cellular contexts. Using nuclear magnetic resonance (NMR) spectroscopy and immunofluorescence analysis we demonstrate that ERK2 scaffolding by hNHE1 occurs by one of three D-domains and by two non-canonical F-sites located in the disordered intracellular tail of hNHE1, mutation of which reduced cellular hNHE1-ERK1/2 co-localization, as well as reduced cellular ERK1/2 activation. Time-resolved NMR spectroscopy revealed that ERK2 phosphorylated the disordered tail of hNHE1 at six sites in vitro, in a distinct temporal order, with the phosphorylation rates at the individual sites being modulated by the docking sites in a distant dependent manner. CONCLUSIONS: This work characterizes a new type of scaffolding complex, which we term a “shuffle complex”, between the disordered hNHE1-tail and ERK2, and provides a molecular mechanism for the important ERK2 scaffolding function of the membrane protein hNHE1, which regulates the phosphorylation of both hNHE1 and ERK2. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12915-016-0252-7) contains supplementary material, which is available to authorized users.
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spelling pubmed-48339482016-04-17 The human Na(+)/H(+) exchanger 1 is a membrane scaffold protein for extracellular signal-regulated kinase 2 Hendus-Altenburger, Ruth Pedraz-Cuesta, Elena Olesen, Christina W. Papaleo, Elena Schnell, Jeff A. Hopper, Jonathan T. S. Robinson, Carol V. Pedersen, Stine F. Kragelund, Birthe B. BMC Biol Research Article BACKGROUND: Extracellular signal-regulated kinase 2 (ERK2) is an S/T kinase with more than 200 known substrates, and with critical roles in regulation of cell growth and differentiation and currently no membrane proteins have been linked to ERK2 scaffolding. METHODS AND RESULTS: Here, we identify the human Na(+)/H(+) exchanger 1 (hNHE1) as a membrane scaffold protein for ERK2 and show direct hNHE1-ERK1/2 interaction in cellular contexts. Using nuclear magnetic resonance (NMR) spectroscopy and immunofluorescence analysis we demonstrate that ERK2 scaffolding by hNHE1 occurs by one of three D-domains and by two non-canonical F-sites located in the disordered intracellular tail of hNHE1, mutation of which reduced cellular hNHE1-ERK1/2 co-localization, as well as reduced cellular ERK1/2 activation. Time-resolved NMR spectroscopy revealed that ERK2 phosphorylated the disordered tail of hNHE1 at six sites in vitro, in a distinct temporal order, with the phosphorylation rates at the individual sites being modulated by the docking sites in a distant dependent manner. CONCLUSIONS: This work characterizes a new type of scaffolding complex, which we term a “shuffle complex”, between the disordered hNHE1-tail and ERK2, and provides a molecular mechanism for the important ERK2 scaffolding function of the membrane protein hNHE1, which regulates the phosphorylation of both hNHE1 and ERK2. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12915-016-0252-7) contains supplementary material, which is available to authorized users. BioMed Central 2016-04-15 /pmc/articles/PMC4833948/ /pubmed/27083547 http://dx.doi.org/10.1186/s12915-016-0252-7 Text en © Hendus-Altenburger et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Hendus-Altenburger, Ruth
Pedraz-Cuesta, Elena
Olesen, Christina W.
Papaleo, Elena
Schnell, Jeff A.
Hopper, Jonathan T. S.
Robinson, Carol V.
Pedersen, Stine F.
Kragelund, Birthe B.
The human Na(+)/H(+) exchanger 1 is a membrane scaffold protein for extracellular signal-regulated kinase 2
title The human Na(+)/H(+) exchanger 1 is a membrane scaffold protein for extracellular signal-regulated kinase 2
title_full The human Na(+)/H(+) exchanger 1 is a membrane scaffold protein for extracellular signal-regulated kinase 2
title_fullStr The human Na(+)/H(+) exchanger 1 is a membrane scaffold protein for extracellular signal-regulated kinase 2
title_full_unstemmed The human Na(+)/H(+) exchanger 1 is a membrane scaffold protein for extracellular signal-regulated kinase 2
title_short The human Na(+)/H(+) exchanger 1 is a membrane scaffold protein for extracellular signal-regulated kinase 2
title_sort human na(+)/h(+) exchanger 1 is a membrane scaffold protein for extracellular signal-regulated kinase 2
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4833948/
https://www.ncbi.nlm.nih.gov/pubmed/27083547
http://dx.doi.org/10.1186/s12915-016-0252-7
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