The human Na+/H+ exchanger 1 is a membrane scaffold protein for extracellular signal-regulated kinase 2
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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 Wilkens; Papaleo, Elena; Schnell, Jeff Alexander; Hopper, Jonathan T. S.; Robinson, Carol V.; Pedersen, Stine Helene Falsig; Kragelund, Birthe Brandt.
I: BMC Biology, Bind 14, 31, 2016.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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T1 - The human Na+/H+ exchanger 1 is a membrane scaffold protein for extracellular signal-regulated kinase 2
AU - Hendus-Altenburger, Ruth
AU - Pedraz Cuesta, Elena
AU - Olesen, Christina Wilkens
AU - Papaleo, Elena
AU - Schnell, Jeff Alexander
AU - Hopper, Jonathan T. S.
AU - Robinson, Carol V.
AU - Pedersen, Stine Helene Falsig
AU - Kragelund, Birthe Brandt
PY - 2016
Y1 - 2016
N2 - 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.
AB - 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.
U2 - 10.1186/s12915-016-0252-7
DO - 10.1186/s12915-016-0252-7
M3 - Journal article
C2 - 27083547
VL - 14
JO - B M C Biology
JF - B M C Biology
SN - 1741-7007
M1 - 31
ER -
ID: 161362945