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 the...
| Main Authors: | , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
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BioMed Central
2016
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| _version_ | 1826258985960865792 |
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| author | Hendus-Altenburger, R Pedraz-Cuesta, E Olesen, C Papaleo, E Schnell, J Hopper, J Robinson, C Pedersen, S Kragelund, B |
| author_facet | Hendus-Altenburger, R Pedraz-Cuesta, E Olesen, C Papaleo, E Schnell, J Hopper, J Robinson, C Pedersen, S Kragelund, B |
| author_sort | Hendus-Altenburger, R |
| collection | OXFORD |
| 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. |
| first_indexed | 2024-03-06T18:42:43Z |
| format | Journal article |
| id | oxford-uuid:0d71af1f-8ffc-4a6b-b689-074c84f65562 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T18:42:43Z |
| publishDate | 2016 |
| publisher | BioMed Central |
| record_format | dspace |
| spelling | oxford-uuid:0d71af1f-8ffc-4a6b-b689-074c84f655622022-03-26T09:40:32ZThe human Na+/H+ exchanger 1 is a membrane scaffold protein for extracellular signal-regulated kinase 2Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:0d71af1f-8ffc-4a6b-b689-074c84f65562EnglishSymplectic Elements at OxfordBioMed Central2016Hendus-Altenburger, RPedraz-Cuesta, EOlesen, CPapaleo, ESchnell, JHopper, JRobinson, CPedersen, SKragelund, BBackground 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. |
| spellingShingle | Hendus-Altenburger, R Pedraz-Cuesta, E Olesen, C Papaleo, E Schnell, J Hopper, J Robinson, C Pedersen, S Kragelund, 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 |
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