The human Na+/H+ exchanger 1 is a membrane scaffold protein for extracellular signal-regulated kinase 2
Tóm tắt
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. 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. 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.
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citation_journal_title=Science; citation_title=Scaffold proteins: hubs for controlling the flow of cellular information; citation_author=MC Good, JG Zalatan, WA Lim; citation_volume=332; citation_publication_date=2011; citation_pages=680-6; citation_doi=10.1126/science.1198701; citation_id=CR1
citation_journal_title=Nat Rev Mol Cell Biol; citation_title=Signalling scaffolds and local organization of cellular behaviour; citation_author=LK Langeberg, JD Scott; citation_volume=16; citation_publication_date=2015; citation_pages=232-44; citation_doi=10.1038/nrm3966; citation_id=CR2
citation_journal_title=J Cell Sci; citation_title=KSR: a MAPK scaffold of the Ras pathway?; citation_author=DK Morrison; citation_volume=114; citation_publication_date=2001; citation_pages=1609-12; citation_id=CR3
citation_journal_title=Mol Cell Biol; citation_title=The JIP group of mitogen-activated protein kinase scaffold proteins; citation_author=J Yasuda, AJ Whitmarsh, J Cavanagh, M Sharma, RJ Davis; citation_volume=19; citation_publication_date=1999; citation_pages=7245-54; citation_doi=10.1128/MCB.19.10.7245; citation_id=CR4
citation_journal_title=Mol Cell Biol; citation_title=IQGAP1 is a scaffold for mitogen-activated protein kinase signaling; citation_author=M Roy, Z Li, DB Sacks; citation_volume=25; citation_publication_date=2005; citation_pages=7940-52; citation_doi=10.1128/MCB.25.18.7940-7952.2005; citation_id=CR5
citation_journal_title=Science; citation_title=Beta-arrestin 2: a receptor-regulated MAPK scaffold for the activation of JNK3; citation_author=PH McDonald, CW Chow, WE Miller, SA Laporte, ME Field, FT Lin; citation_volume=290; citation_publication_date=2000; citation_pages=1574-7; citation_doi=10.1126/science.290.5496.1574; citation_id=CR6
citation_journal_title=Sci Signal; citation_title=Specificity of linear motifs that bind to a common mitogen-activated protein kinase docking groove; citation_author=A Garai, A Zeke, G Gogl, I Toro, F Fordos, H Blankenburg; citation_volume=5; citation_publication_date=2012; citation_pages=ra74; citation_doi=10.1126/scisignal.2003004; citation_id=CR7
citation_journal_title=Pharmacol Res; citation_title=ERK1/2 MAP kinases: structure, function, and regulation; citation_author=R Roskoski; citation_volume=66; citation_publication_date=2012; citation_pages=105-43; citation_doi=10.1016/j.phrs.2012.04.005; citation_id=CR8
citation_journal_title=PLoS Comput Biol; citation_title=Computational prediction and experimental verification of new MAP kinase docking sites and substrates including Gli transcription factors; citation_author=TC Whisenant, DT Ho, RW Benz, JS Rogers, RM Kaake, EA Gordon; citation_volume=6; citation_publication_date=2010; citation_pages=e1000908; citation_doi=10.1371/journal.pcbi.1000908; citation_id=CR9
citation_journal_title=Nat Cell Biol; citation_title=A conserved docking motif in MAP kinases common to substrates, activators and regulators; citation_author=T Tanoue, M Adachi, T Moriguchi, E Nishida; citation_volume=2; citation_publication_date=2000; citation_pages=110-6; citation_doi=10.1038/35000065; citation_id=CR10
citation_journal_title=Mol Cell; citation_title=Docking motif interactions in MAP kinases revealed by hydrogen exchange mass spectrometry; citation_author=T Lee, AN Hoofnagle, Y Kabuyama, J Stroud, X Min, EJ Goldsmith; citation_volume=14; citation_publication_date=2004; citation_pages=43-55; citation_doi=10.1016/S1097-2765(04)00161-3; citation_id=CR11
citation_journal_title=Genes Dev; citation_title=Multiple docking sites on substrate proteins form a modular system that mediates recognition by ERK MAP kinase; citation_author=D Jacobs, D Glossip, H Xing, AJ Muslin, K Kornfeld; citation_volume=13; citation_publication_date=1999; citation_pages=163-75; citation_doi=10.1101/gad.13.2.163; citation_id=CR12
citation_journal_title=J Biol Chem; citation_title=Substrate discrimination among mitogen-activated protein kinases through distinct docking sequence motifs; citation_author=DL Sheridan, Y Kong, SA Parker, KN Dalby, BE Turk; citation_volume=283; citation_publication_date=2008; citation_pages=19511-20; citation_doi=10.1074/jbc.M801074200; citation_id=CR13
citation_journal_title=Genetics; citation_title=Gain-of-function mutations in the Caenorhabditis elegans lin-1 ETS gene identify a C-terminal regulatory domain phosphorylated by ERK MAP kinase; citation_author=D Jacobs, GJ Beitel, SG Clark, HR Horvitz, K Kornfeld; citation_volume=149; citation_publication_date=1998; citation_pages=1809-22; citation_id=CR14
citation_journal_title=Nat Cell Biol; citation_title=Molecular interpretation of ERK signal duration by immediate early gene products; citation_author=LO Murphy, S Smith, RH Chen, DC Fingar, J Blenis; citation_volume=4; citation_publication_date=2002; citation_pages=556-64; citation_id=CR15
citation_journal_title=Nat Struct Mol Biol; citation_title=Phosphoproteomics reveals new ERK MAP kinase targets and links ERK to nucleoporin-mediated nuclear transport; citation_author=H Kosako, N Yamaguchi, C Aranami, M Ushiyama, S Kose, N Imamoto; citation_volume=16; citation_publication_date=2009; citation_pages=1026-35; citation_doi=10.1038/nsmb.1656; citation_id=CR16
citation_journal_title=J Biol Chem; citation_title=Evidence for existence of a nuclear pore complex-mediated, cytosol-independent pathway of nuclear translocation of ERK MAP kinase in permeabilized cells; citation_author=Y Matsubayashi, M Fukuda, E Nishida; citation_volume=276; citation_publication_date=2001; citation_pages=41755-60; citation_doi=10.1074/jbc.M106012200; citation_id=CR17
citation_journal_title=Nat Commun; citation_title=Structure of ERK2 bound to PEA-15 reveals a mechanism for rapid release of activated MAPK; citation_author=PD Mace, Y Wallez, MF Egger, MK Dobaczewska, H Robinson, EB Pasquale; citation_volume=4; citation_publication_date=2013; citation_pages=1681; citation_doi=10.1038/ncomms2687; citation_id=CR18
citation_journal_title=Pflugers Arch; citation_title=The Na+/H+ exchanger NHE1 in stress-induced signal transduction: implications for cell proliferation and cell death; citation_author=SF Pedersen; citation_volume=452; citation_publication_date=2006; citation_pages=249-59; citation_doi=10.1007/s00424-006-0044-y; citation_id=CR19
citation_journal_title=Curr Pharm Des; citation_title=Physiology, pharmacology and pathophysiology of the pH regulatory transport proteins NHE1 and NBCn1: similarities, differences, and implications for cancer therapy; citation_author=E Boedtkjer, L Bunch, SF Pedersen; citation_volume=18; citation_publication_date=2012; citation_pages=1345-71; citation_doi=10.2174/138161212799504830; citation_id=CR20
citation_journal_title=J Biol Chem; citation_title=Multiple receptors coupled to adenylate cyclase regulate Na-H exchange independent of cAMP; citation_author=MB Ganz, JA Pachter, DL Barber; citation_volume=265; citation_publication_date=1990; citation_pages=8989-92; citation_id=CR21
citation_journal_title=Cancer Lett; citation_title=The Na+/H+ exchanger NHE1, but not the Na+, HCO3-cotransporter NBCn1, regulates motility of MCF7 breast cancer cells expressing constitutively active ErbB2; citation_author=G Lauritzen, CM Stock, J Lemaire, SF Lund, MF Jensen, B Damsgaard; citation_volume=317; citation_publication_date=2012; citation_pages=172-83; citation_doi=10.1016/j.canlet.2011.11.023; citation_id=CR22
citation_journal_title=J Biol Chem; citation_title=Growth factor activation and “H+-sensing” of the Na+/H+ exchanger isoform 1 (NHE1). Evidence for an additional mechanism not requiring direct phosphorylation; citation_author=S Wakabayashi, B Bertrand, M Shigekawa, P Fafournoux, J Pouyssegur; citation_volume=269; citation_publication_date=1994; citation_pages=5583-8; citation_id=CR23
citation_journal_title=J Biol Chem; citation_title=Stimulation of the plasma membrane Na+/H+ exchanger NHE1 by sustained intracellular acidosis. Evidence for a novel mechanism mediated by the ERK pathway; citation_author=RS Haworth, C McCann, AK Snabaitis, NA Roberts, M Avkiran; citation_volume=278; citation_publication_date=2003; citation_pages=31676-84; citation_doi=10.1074/jbc.M304400200; citation_id=CR24
citation_journal_title=J Biol Chem; citation_title=Mitogen-activated protein kinase-dependent activation of the Na+/H+ exchanger is mediated through phosphorylation of amino acids Ser770 and Ser771; citation_author=ME Malo, L Li, L Fliegel; citation_volume=282; citation_publication_date=2007; citation_pages=6292-9; citation_doi=10.1074/jbc.M611073200; citation_id=CR25
citation_journal_title=Cell Physiol Biochem; citation_title=The Na+/H+ exchanger, NHE1, differentially regulates mitogen-activated protein kinase subfamilies after osmotic shrinkage in Ehrlich Lettre Ascites cells; citation_author=SF Pedersen, BV Darborg, M Rasmussen, J Nylandsted, EK Hoffmann; citation_volume=20; citation_publication_date=2007; citation_pages=735-50; citation_doi=10.1159/000110434; citation_id=CR26
citation_journal_title=J Biol Chem; citation_title=p90(RSK) is a serum-stimulated Na+/H+ exchanger isoform-1 kinase. Regulatory phosphorylation of serine 703 of Na+/H+ exchanger isoform-1; citation_author=E Takahashi, J Abe, B Gallis, R Aebersold, DJ Spring, EG Krebs; citation_volume=274; citation_publication_date=1999; citation_pages=20206-14; citation_doi=10.1074/jbc.274.29.20206; citation_id=CR27
citation_journal_title=Arch Biochem Biophys; citation_title=Regulation of mitogen-activated protein kinase pathways by the plasma membrane Na+/H+ exchanger, NHE1; citation_author=SF Pedersen, BV Darborg, ML Rentsch, M Rasmussen; citation_volume=462; citation_publication_date=2007; citation_pages=195-201; citation_doi=10.1016/j.abb.2006.12.001; citation_id=CR28
citation_journal_title=Anal Chem; citation_title=Open tubular immobilized metal ion affinity chromatography combined with MALDI MS and MS/MS for identification of protein phosphorylation sites; citation_author=H Liu, J Stupak, J Zheng, BO Keller, BJ Brix, L Fliegel; citation_volume=76; citation_publication_date=2004; citation_pages=4223-32; citation_doi=10.1021/ac035231d; citation_id=CR29
citation_journal_title=J Pharmacol Exp Ther; citation_title=Antihypertrophic effect of Na+/H+ exchanger isoform 1 inhibition is mediated by reduced mitogen-activated protein kinase activation secondary to improved mitochondrial integrity and decreased generation of mitochondrial-derived reactive oxygen species; citation_author=S Javadov, D Baetz, V Rajapurohitam, A Zeidan, LA Kirshenbaum, M Karmazyn; citation_volume=317; citation_publication_date=2006; citation_pages=1036-43; citation_doi=10.1124/jpet.105.100107; citation_id=CR30
citation_journal_title=J Biol Chem; citation_title=ERK is regulated by sodium-proton exchanger in rat aortic vascular smooth muscle cells; citation_author=YV Mukhin, MN Garnovskaya, ME Ullian, JR Raymond; citation_volume=279; citation_publication_date=2004; citation_pages=1845-52; citation_doi=10.1074/jbc.M304907200; citation_id=CR31
citation_journal_title=Pflugers Arch; citation_title=Roles of Na+/H+ exchange in regulation of p38 mitogen-activated protein kinase activity and cell death after chemical anoxia in NIH3T3 fibroblasts; citation_author=ML Rentsch, CG Ossum, EK Hoffmann, SF Pedersen; citation_volume=454; citation_publication_date=2007; citation_pages=649-62; citation_doi=10.1007/s00424-007-0233-3; citation_id=CR32
citation_journal_title=Nat Methods; citation_title=A human MAP kinase interactome; citation_author=S Bandyopadhyay, CY Chiang, J Srivastava, M Gersten, S White, R Bell; citation_volume=7; citation_publication_date=2010; citation_pages=801-5; citation_doi=10.1038/nmeth.1506; citation_id=CR33
citation_journal_title=J Biol Chem; citation_title=B-Raf associates with and activates the NHE1 isoform of the Na+/H+ exchanger; citation_author=P Karki, X Li, D Schrama, L Fliegel; citation_volume=286; citation_publication_date=2011; citation_pages=13096-105; citation_doi=10.1074/jbc.M110.165134; citation_id=CR34
citation_journal_title=Biochemistry; citation_title=The intracellular distal tail of the Na+/H+ exchanger NHE1 is intrinsically disordered: implications for NHE1 trafficking; citation_author=AB Norholm, R Hendus-Altenburger, G Bjerre, M Kjaergaard, SF Pedersen, BB Kragelund; citation_volume=50; citation_publication_date=2011; citation_pages=3469-80; citation_doi=10.1021/bi1019989; citation_id=CR35
citation_journal_title=Curr Top Membr; citation_title=Structural dynamics and regulation of the mammalian SLC9A family of Na+/H+ exchangers; citation_author=R Hendus-Altenburger, BB Kragelund, SF Pedersen; citation_volume=73; citation_publication_date=2014; citation_pages=69-148; citation_doi=10.1016/B978-0-12-800223-0.00002-5; citation_id=CR36
citation_journal_title=J Mol Biol; citation_title=Prediction and functional analysis of native disorder in proteins from the three kingdoms of life; citation_author=JJ Ward, JS Sodhi, LJ McGuffin, BF Buxton, DT Jones; citation_volume=337; citation_publication_date=2004; citation_pages=635-45; citation_doi=10.1016/j.jmb.2004.02.002; citation_id=CR37
citation_journal_title=J Mol Biol; citation_title=Intrinsic disorder in cell-signaling and cancer-associated proteins; citation_author=LM Iakoucheva, CJ Brown, JD Lawson, Z Obradovic, AK Dunker; citation_volume=323; citation_publication_date=2002; citation_pages=573-84; citation_doi=10.1016/S0022-2836(02)00969-5; citation_id=CR38
citation_journal_title=Prog Biophys Mol Biol; citation_title=Intrinsic disorder in scaffold proteins: getting more from less; citation_author=MS Cortese, VN Uversky, DA Keith; citation_volume=98; citation_publication_date=2008; citation_pages=85-106; citation_doi=10.1016/j.pbiomolbio.2008.05.007; citation_id=CR39
citation_journal_title=Proc Natl Acad Sci U S A; citation_title=Structure and dynamics of the MKK7-JNK signaling complex; citation_author=J Kragelj, A Palencia, MH Nanao, D Maurin, G Bouvignies, M Blackledge; citation_volume=112; citation_publication_date=2015; citation_pages=3409-14; citation_doi=10.1073/pnas.1419528112; citation_id=CR40
citation_journal_title=Am J Physiol; citation_title=Impaired cell volume regulation in Na(+)-H+ exchange-deficient mutants; citation_author=D Rotin, S Grinstein; citation_volume=257; citation_publication_date=1989; citation_pages=C1158-65; citation_id=CR41
citation_journal_title=J Biol; citation_title=Distinct functions for ERKs?; citation_author=AC Lloyd; citation_volume=5; citation_publication_date=2006; citation_pages=13; citation_doi=10.1186/jbiol46; citation_id=CR42
citation_journal_title=J Biol Chem; citation_title=Regulation and properties of extracellular signal-regulated protein kinases 1 and 2 in vitro; citation_author=DJ Robbins, E Zhen, H Owaki, CA Vanderbilt, D Ebert, TD Geppert; citation_volume=268; citation_publication_date=1993; citation_pages=5097-106; citation_id=CR43
citation_journal_title=J Biol Chem; citation_title=The activity of the extracellular signal-regulated kinase 2 is regulated by differential phosphorylation in the activation loop; citation_author=B Zhou, ZY Zhang; citation_volume=277; citation_publication_date=2002; citation_pages=13889-99; citation_doi=10.1074/jbc.M200377200; citation_id=CR44
citation_journal_title=EMBO J; citation_title=Differential targeting of MAP kinases to the ETS-domain transcription factor Elk-1; citation_author=SH Yang, AJ Whitmarsh, RJ Davis, AD Sharrocks; citation_volume=17; citation_publication_date=1998; citation_pages=1740-9; citation_doi=10.1093/emboj/17.6.1740; citation_id=CR45
citation_journal_title=Anal Chem; citation_title=Bayesian deconvolution of mass and ion mobility spectra: from binary interactions to polydisperse ensembles; citation_author=MT Marty, AJ Baldwin, EG Marklund, GK Hochberg, JL Benesch, CV Robinson; citation_volume=87; citation_publication_date=2015; citation_pages=4370-6; citation_doi=10.1021/acs.analchem.5b00140; citation_id=CR46
citation_journal_title=Biochemistry; citation_title=Activated ERK2 is a monomer in vitro with or without divalent cations and when complexed to the cytoplasmic scaffold PEA-15; citation_author=TS Kaoud, AK Devkota, R Harris, MS Rana, O Abramczyk, M Warthaka; citation_volume=50; citation_publication_date=2011; citation_pages=4568-78; citation_doi=10.1021/bi200202y; citation_id=CR47
citation_journal_title=Nat Commun; citation_title=Quantitative maps of protein phosphorylation sites across 14 different rat organs and tissues; citation_author=A Lundby, A Secher, K Lage, NB Nordsborg, A Dmytriyev, C Lundby; citation_volume=3; citation_publication_date=2012; citation_pages=876; citation_doi=10.1038/ncomms1871; citation_id=CR48
citation_journal_title=J Biomol NMR; citation_title=Cell signaling, post-translational protein modifications and NMR spectroscopy; citation_author=FX Theillet, C Smet-Nocca, S Liokatis, R Thongwichian, J Kosten, MK Yoon; citation_volume=54; citation_publication_date=2012; citation_pages=217-36; citation_doi=10.1007/s10858-012-9674-x; citation_id=CR49
citation_journal_title=J Biomol NMR; citation_title=Unraveling a phosphorylation event in a folded protein by NMR spectroscopy: phosphorylation of the Pin1 WW domain by PKA; citation_author=C Smet-Nocca, H Launay, JM Wieruszeski, G Lippens, I Landrieu; citation_volume=55; citation_publication_date=2013; citation_pages=323-37; citation_doi=10.1007/s10858-013-9716-z; citation_id=CR50
citation_journal_title=J Cell Sci; citation_title=Regulation of the formation of tumor cell pseudopodia by the Na+/H+ exchanger NHE1; citation_author=A Lagana, J Vadnais, PU Le, TN Nguyen, R Laprade, IR Nabi; citation_volume=113; citation_issue=Pt 20; citation_publication_date=2000; citation_pages=3649-62; citation_id=CR51
citation_journal_title=J Cell Biol; citation_title=The Na+/H+ exchanger NHE1 is required for directional migration stimulated via PDGFR-alpha in the primary cilium; citation_author=L Schneider, CM Stock, P Dieterich, BH Jensen, LB Pedersen, P Satir; citation_volume=185; citation_publication_date=2009; citation_pages=163-76; citation_doi=10.1083/jcb.200806019; citation_id=CR52
citation_journal_title=Am J Physiol Cell Physiol; citation_title=Intracellular pH gradients in migrating cells; citation_author=C Martin, SF Pedersen, A Schwab, C Stock; citation_volume=300; citation_publication_date=2011; citation_pages=C490-5; citation_doi=10.1152/ajpcell.00280.2010; citation_id=CR53
Pedraz-Cuesta E, Fredsted J, Jensen HH, Bornebusch A, Nejsum LN, Kragelund BB, et al. Prolactin signaling stimulates invasion via the Na+/H+ exchanger NHE1 in T47D human breast cancer cells. Mol Endocrinol. 2015; in revision
citation_journal_title=Cell; citation_title=Global, in vivo, and site-specific phosphorylation dynamics in signaling networks; citation_author=JV Olsen, B Blagoev, F Gnad, B Macek, C Kumar, P Mortensen; citation_volume=127; citation_publication_date=2006; citation_pages=635-48; citation_doi=10.1016/j.cell.2006.09.026; citation_id=CR55
citation_journal_title=Chem Soc Rev; citation_title=Multitude of binding modes attainable by intrinsically disordered proteins: a portrait gallery of disorder-based complexes; citation_author=VN Uversky; citation_volume=40; citation_publication_date=2011; citation_pages=1623-34; citation_doi=10.1039/C0CS00057D; citation_id=CR56
citation_journal_title=Science; citation_title=The Ste5 scaffold allosterically modulates signaling output of the yeast mating pathway; citation_author=RP Bhattacharyya, A Remenyi, MC Good, CJ Bashor, AM Falick, WA Lim; citation_volume=311; citation_publication_date=2006; citation_pages=822-6; citation_doi=10.1126/science.1120941; citation_id=CR57
citation_journal_title=Nat Chem Biol; citation_title=Intrinsic disorder mediates the diverse regulatory functions of the Cdk inhibitor p21; citation_author=Y Wang, JC Fisher, R Mathew, L Ou, S Otieno, J Sublet; citation_volume=7; citation_publication_date=2011; citation_pages=214-21; citation_doi=10.1038/nchembio.536; citation_id=CR58
citation_journal_title=Structure; citation_title=Structure/function implications in a dynamic complex of the intrinsically disordered Sic1 with the Cdc4 subunit of an SCF ubiquitin ligase; citation_author=T Mittag, J Marsh, A Grishaev, S Orlicky, H Lin, F Sicheri; citation_volume=18; citation_publication_date=2010; citation_pages=494-506; citation_doi=10.1016/j.str.2010.01.020; citation_id=CR59
citation_journal_title=Proc Natl Acad Sci U S A; citation_title=Composite low affinity interactions dictate recognition of the cyclin-dependent kinase inhibitor Sic1 by the SCFCdc4 ubiquitin ligase; citation_author=X Tang, S Orlicky, T Mittag, V Csizmok, T Pawson, JD Forman-Kay; citation_volume=109; citation_publication_date=2012; citation_pages=3287-92; citation_doi=10.1073/pnas.1116455109; citation_id=CR60
citation_journal_title=Philos Trans R Soc Lond B Biol Sci; citation_title=Self-assembly of biological macromolecules; citation_author=RN Perham; citation_volume=272; citation_publication_date=1975; citation_pages=123-36; citation_doi=10.1098/rstb.1975.0075; citation_id=CR61
citation_journal_title=Biochem J; citation_title=Docking sites on mitogen-activated protein kinase (MAPK) kinases, MAPK phosphatases and the Elk-1 transcription factor compete for MAPK binding and are crucial for enzymic activity; citation_author=AJ Bardwell, M Abdollahi, L Bardwell; citation_volume=370; citation_publication_date=2003; citation_pages=1077-85; citation_doi=10.1042/bj20021806; citation_id=CR62
citation_journal_title=Biochemistry; citation_title=Dimerization is crucial for the function of the Na+/H+ exchanger NHE1; citation_author=T Hisamitsu, YB Ammar, TY Nakamura, S Wakabayashi; citation_volume=45; citation_publication_date=2006; citation_pages=13346-55; citation_doi=10.1021/bi0608616; citation_id=CR63
citation_journal_title=PLoS One; citation_title=Interface analysis of the complex between ERK2 and PTP-SL; citation_author=MC Balasu, LN Spiridon, S Miron, CT Craescu, AJ Scheidig, AJ Petrescu; citation_volume=4; citation_publication_date=2009; citation_pages=e5432; citation_doi=10.1371/journal.pone.0005432; citation_id=CR64
citation_journal_title=Proc Natl Acad Sci U S A; citation_title=A quantitative atlas of mitotic phosphorylation; citation_author=N Dephoure, C Zhou, J Villen, SA Beausoleil, CE Bakalarski, SJ Elledge; citation_volume=105; citation_publication_date=2008; citation_pages=10762-7; citation_doi=10.1073/pnas.0805139105; citation_id=CR65
citation_journal_title=Mol Cell Biol; citation_title=Trophic factor withdrawal: p38 mitogen-activated protein kinase activates NHE1, which induces intracellular alkalinization; citation_author=AR Khaled, AN Moor, A Li, K Kim, DK Ferris, K Muegge; citation_volume=21; citation_publication_date=2001; citation_pages=7545-57; citation_doi=10.1128/MCB.21.22.7545-7557.2001; citation_id=CR66
citation_journal_title=Am J Physiol Cell Physiol; citation_title=Apoptosis-induced alkalinization by the Na+/H+ exchanger isoform 1 is mediated through phosphorylation of amino acids Ser726 and Ser729; citation_author=AL Grenier, K Abu-Ihweij, G Zhang, SM Ruppert, R Boohaker, ER Slepkov; citation_volume=295; citation_publication_date=2008; citation_pages=C883-96; citation_doi=10.1152/ajpcell.00574.2007; citation_id=CR67
citation_journal_title=Am J Physiol Renal Physiol; citation_title=Acidosis-mediated regulation of the NHE1 isoform of the Na+/H+ exchanger in renal cells; citation_author=A Odunewu, L Fliegel; citation_volume=305; citation_publication_date=2013; citation_pages=F370-81; citation_doi=10.1152/ajprenal.00598.2012; citation_id=CR68
citation_journal_title=PLoS One; citation_title=Phosphorylation and activation of the plasma membrane Na+/H+ exchanger (NHE1) during osmotic cell shrinkage; citation_author=RR Rigor, C Damoc, BS Phinney, PM Cala; citation_volume=6; citation_publication_date=2011; citation_pages=e29210; citation_doi=10.1371/journal.pone.0029210; citation_id=CR69
citation_journal_title=Nat Struct Mol Biol; citation_title=In situ observation of protein phosphorylation by high-resolution NMR spectroscopy; citation_author=P Selenko, DP Frueh, SJ Elsaesser, W Haas, SP Gygi, G Wagner; citation_volume=15; citation_publication_date=2008; citation_pages=321-9; citation_doi=10.1038/nsmb.1395; citation_id=CR70
citation_journal_title=Protein Sci; citation_title=Temperature-dependent structural changes in intrinsically disordered proteins: formation of alpha-helices or loss of polyproline II?; citation_author=M Kjaergaard, AB Norholm, R Hendus-Altenburger, SF Pedersen, FM Poulsen, BB Kragelund; citation_volume=19; citation_publication_date=2010; citation_pages=1555-64; citation_doi=10.1002/pro.435; citation_id=CR71
citation_journal_title=Nucleic Acids Res; citation_title=Scansite 2.0: proteome-wide prediction of cell signaling interactions using short sequence motifs; citation_author=JC Obenauer, LC Cantley, MB Yaffe; citation_volume=31; citation_publication_date=2003; citation_pages=3635-41; citation_doi=10.1093/nar/gkg584; citation_id=CR72
citation_journal_title=Mol Syst Biol; citation_title=Fast, scalable generation of high-quality protein multiple sequence alignments using Clustal Omega; citation_author=F Sievers, A Wilm, D Dineen, TJ Gibson, K Karplus, W Li; citation_volume=7; citation_publication_date=2011; citation_pages=539; citation_doi=10.1038/msb.2011.75; citation_id=CR73
citation_journal_title=Biochim Biophys Acta; citation_title=PONDR-FIT: a meta-predictor of intrinsically disordered amino acids; citation_author=B Xue, RL Dunbrack, RW Williams, AK Dunker, VN Uversky; citation_volume=1804; citation_publication_date=2010; citation_pages=996-1010; citation_doi=10.1016/j.bbapap.2010.01.011; citation_id=CR74
citation_journal_title=Bioinformatics; citation_title=The DISOPRED server for the prediction of protein disorder; citation_author=JJ Ward, LJ McGuffin, K Bryson, BF Buxton, DT Jones; citation_volume=20; citation_publication_date=2004; citation_pages=2138-9; citation_doi=10.1093/bioinformatics/bth195; citation_id=CR75
citation_journal_title=J Biomol NMR; citation_title=NMRPipe: a multidimensional spectral processing system based on UNIX pipes; citation_author=F Delaglio, S Grzesiek, GW Vuister, G Zhu, J Pfeifer, A Bax; citation_volume=6; citation_publication_date=1995; citation_pages=277-93; citation_doi=10.1007/BF00197809; citation_id=CR76
citation_journal_title=Proteins; citation_title=The CCPN data model for NMR spectroscopy: development of a software pipeline; citation_author=WF Vranken, W Boucher, TJ Stevens, RH Fogh, A Pajon, M Llinas; citation_volume=59; citation_publication_date=2005; citation_pages=687-96; citation_doi=10.1002/prot.20449; citation_id=CR77
citation_journal_title=Anal Chem; citation_title=A tandem mass spectrometer for improved transmission and analysis of large macromolecular assemblies; citation_author=F Sobott, H Hernandez, MG McCammon, MA Tito, CV Robinson; citation_volume=74; citation_publication_date=2002; citation_pages=1402-7; citation_doi=10.1021/ac0110552; citation_id=CR78
citation_journal_title=Nat Protoc; citation_title=Determining the stoichiometry and interactions of macromolecular assemblies from mass spectrometry; citation_author=H Hernandez, CV Robinson; citation_volume=2; citation_publication_date=2007; citation_pages=715-26; citation_doi=10.1038/nprot.2007.73; citation_id=CR79
citation_journal_title=Acta Crystallogr D Biol Crystallogr; citation_title=Protein-peptide complex crystallization: a case study on the ERK2 mitogen-activated protein kinase; citation_author=G Gogl, I Toro, A Remenyi; citation_volume=69; citation_publication_date=2013; citation_pages=486-9; citation_doi=10.1107/S0907444912051062; citation_id=CR80
citation_journal_title=Methods Enzymol; citation_title=Modeller: generation and refinement of homology-based protein structure models; citation_author=A Fiser, A Sali; citation_volume=374; citation_publication_date=2003; citation_pages=461-91; citation_doi=10.1016/S0076-6879(03)74020-8; citation_id=CR81
citation_journal_title=J Biol Chem; citation_title=NHE1 inhibition by amiloride- and benzoylguanidine-type compounds. Inhibitor binding loci deduced from chimeras of NHE1 homologues with endogenous differences in inhibitor sensitivity; citation_author=SF Pedersen, SA King, EB Nygaard, RR Rigor, PM Cala; citation_volume=282; citation_publication_date=2007; citation_pages=19716-27; citation_doi=10.1074/jbc.M701637200; citation_id=CR82