Cargando…
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...
Autores principales: | , , , , , , , , |
---|---|
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 |
_version_ | 1782427415592566784 |
---|---|
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. |
format | Online Article Text |
id | pubmed-4833948 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
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 |
work_keys_str_mv | AT hendusaltenburgerruth thehumannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT pedrazcuestaelena thehumannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT olesenchristinaw thehumannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT papaleoelena thehumannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT schnelljeffa thehumannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT hopperjonathants thehumannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT robinsoncarolv thehumannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT pedersenstinef thehumannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT kragelundbirtheb thehumannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT hendusaltenburgerruth humannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT pedrazcuestaelena humannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT olesenchristinaw humannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT papaleoelena humannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT schnelljeffa humannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT hopperjonathants humannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT robinsoncarolv humannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT pedersenstinef humannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 AT kragelundbirtheb humannahexchanger1isamembranescaffoldproteinforextracellularsignalregulatedkinase2 |