CD95 pp 95-102 | Cite as

CD95-Mediated Proton Regulation

  • Auréa Cophignon
  • Mallorie Poët
  • Michael Monet
  • Michel Tauc
  • Laurent Counillon
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1557)

Abstract

The Na+/H+ exchanger NHE1 is at the crossroads of a large diversity of signaling pathways, whose activation modifies the cooperative response of the transporter to intracellular H+ ions. Here we show how the activation of the Na+/H+ exchanger NHE1 by the cleaved ligand of CD95 can be measured. We demonstrate two different methods designed to set intracellular pH at precise values. Then we show how these can be coupled to fast kinetics of lithium transport, which will enable to measure the NHE1 activity like for an enzyme, because they will yield rates of transport.

Key words

Ion transport Kinetic measurements Intracellular acidification CD95 signaling 

References

  1. 1.
    Kischkel FC et al (1995) Cytotoxicity-dependent APO-1 (Fas/CD95)-associated proteins form a death-inducing signaling complex (DISC) with the receptor. EMBO J 14(22):5579–5588PubMedPubMedCentralGoogle Scholar
  2. 2.
    Cheng J et al (1994) Protection from Fas-mediated apoptosis by a soluble form of the Fas molecule. Science 263(5154):1759–1762CrossRefPubMedGoogle Scholar
  3. 3.
    Tauzin S et al (2011) The naturally processed CD95L elicits a c-Yes/calcium/PI3K-driven cell migration pathway. PLoS Biol 9(6):e1001090CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Counillon L, Pouysségur J (2000) The expanding family of eucaryotic Na+/H+ exchangers. J Biol Chem 275:1–4CrossRefPubMedGoogle Scholar
  5. 5.
    Counillon L, Bouret Y, Marchiq I, Pouysségur J (2016) Na+/H+ antiporter (NHE1) and lactate/H+ symporters (MCTs) in pH homeostasis and cancer metabolism. Biochim Biophys Acta 1863(10):2465–2480, pii: S0167-4889(16)30043-XCrossRefPubMedGoogle Scholar
  6. 6.
    Pouysségur J et al (1984) A specific mutation abolishing Na+/H+ antiport activity in hamster fibroblasts precludes growth at neutral and acidic pH. Proc Natl Acad Sci U S A 81(15):4833–4837CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Pedersen SF et al (2007) Regulation of mitogen-activated protein kinase pathways by the plasma membrane Na+/H+ exchanger, NHE1. Arch Biochem Biophys 462(2):195–201CrossRefPubMedGoogle Scholar
  8. 8.
    Lacroix J et al (2004) Mechanism for the activation of the Na/H exchanger NHE-1 by cytoplasmic acidification and mitogens. EMBO Rep 5(1):91–96CrossRefPubMedGoogle Scholar
  9. 9.
    Frantz C et al (2007) Positive feedback between Cdc42 activity and H+ efflux by the Na-H exchanger NHE1 for polarity of migrating cells. J Cell Biol 179(3):403–410CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Schneider L et al (2009) The Na+/H+ exchanger NHE1 is required for directional migration stimulated via PDGFR-alpha in the primary cilium. J Cell Biol 185(1):163–176CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Stock C et al (2005) Migration of human melanoma cells depends on extracellular pH and NHE. J Physiol 567(Pt 1):225–238CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Martin C et al (2011) Intracellular pH gradients in migrating cells. Am J Physiol Cell Physiol 300(3):C490–C495CrossRefPubMedGoogle Scholar
  13. 13.
    Stock C et al (2007) pH nanoenvironnement at the surface of single melanoma cells. Cell Physiol Biochem 20(5):679–686CrossRefPubMedGoogle Scholar
  14. 14.
    Lauritzen G et al (2012) The NHE1, but not NBCn1, regulates motility of MCF7 breast cancer cells expressing constitutively active ErbB2. Cancer Lett 317(2):172–183CrossRefPubMedGoogle Scholar
  15. 15.
    Brisson L et al (2013) NaV1.5 Na+ channels allosterically regulate the NHE-1 exchanger and promote the activity of breast cancer cell invadopodia. J Cell Sci 126(Pt 21):4835–4842CrossRefPubMedGoogle Scholar
  16. 16.
    Monet M et al (2016) The cleaved FAS ligand activates the Na+/H+ exchanger NHE1 through Akt/ROCK1 to stimulate cell motility. Sci Rep 6:28008CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Paris S, Pouyssegur J (1984) Growth factors activate the Na+/H+ antiporter in quiescent fibroblasts by increasing its affinity for intracellular H+. J Biol Chem 259:10989–10994PubMedGoogle Scholar
  18. 18.
    Milosavljevic N et al (2010) Nongenomic effects of cisplatin: acute inhibition of mechanosensitive transporters and channels without actin remodeling. Cancer Res 70(19):7514–7522CrossRefPubMedGoogle Scholar
  19. 19.
    Boron W, de Veer P (1976) Intracellular pH transients in squid giant axons caused by CO2, NH3, and metabolic inhibitors. J Gen Physiol 67(1):91–112CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Auréa Cophignon
    • 1
    • 2
  • Mallorie Poët
    • 1
    • 2
  • Michael Monet
    • 1
    • 2
    • 3
  • Michel Tauc
    • 1
    • 2
  • Laurent Counillon
    • 1
    • 2
  1. 1.LP2M UMR 7370 Faculté de MédecineUniversité Nice-Sophia AntipolisNiceFrance
  2. 2.Laboratories of Excellence, Ion Channels Sciences and TherapeuticsUniversité LilleVilleneuve d’AscqFrance
  3. 3.LPCM EA4667Université de Picardie Jules VerneAmiensFrance

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