Abstract
The prototypical death receptor Fas (also known as CD95 or Apo-1) plays an essential role in the maintenance of lymphocyte homeostasis. Propagation of cell death through Fas relies on the formation of a multiprotein complex at the receptor level known as the death-inducing signaling complex (DISC). Here, we describe an immunoprecipitation-based protocol to study DISC assembly in activated human T lymphocytes. This procedure is a useful tool to visualize proteins associated with Fas.
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Krammer PH, Arnold R, Lavrik IN (2007) Life and death in peripheral T cells. Nat Rev Immunol 7(7):532–542
Tibbetts MD, Zheng L, Lenardo MJ (2003) The death effector domain protein family: regulators of cellular homeostasis. Nat Immunol 4(5):404–409
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):5579–5588
Peter ME, Krammer PH (2003) The CD95(APO-1/Fas) DISC and beyond. Cell Death Differ 10(1):26–35
Bidere N, Su HC, Lenardo MJ (2006) Genetic disorders of programmed cell death in the immune system. Annu Rev Immunol 24: 321–352
Thome M et al (1997) Viral FLICE-inhibitory proteins (FLIPs) prevent apoptosis induced by death receptors. Nature 386(6624):517–521
Wang J, Chun HJ, Wong W, Spencer DM, Lenardo MJ (2001) Caspase-10 is an initiator caspase in death receptor signaling. Proc Natl Acad Sci U S A 98(24):13884–13888
Stanger BZ, Leder P, Lee TH, Kim E, Seed B (1995) RIP: a novel protein containing a death domain that interacts with Fas/APO-1 (CD95) in yeast and causes cell death. Cell 81(4): 513–523
Lavrik IN, Golks A, Baumann S, Krammer PH (2006) Caspase-2 is activated at the CD95 death-inducing signaling complex in the course of CD95-induced apoptosis. Blood 108(2): 559–565
Snow AL et al (2009) Restimulation-induced apoptosis of T cells is impaired in patients with X-linked lymphoproliferative disease caused by SAP deficiency. J Clin Invest 119(10): 2976–2989
Bonifacino JS, Dell’Angelica EC, Springer TA (2001) Immunoprecipitation. Curr Protoc Mol Biol Chapter 3:Unit 10 16.
Acknowledgements
This work was supported by grants from ANR JCJC, Fondation de France, Association pour la Recherche contre le Cancer, and Ligue Nationale contre le Cancer.
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Cordeiro, N., Bidère, N. (2013). Visualization of Fas-Mediated Death-Inducing Signaling Complex Formation by Immunoprecipitation. In: Snow, A., Lenardo, M. (eds) Immune Homeostasis. Methods in Molecular Biology, vol 979. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-290-2_5
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DOI: https://doi.org/10.1007/978-1-62703-290-2_5
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-289-6
Online ISBN: 978-1-62703-290-2
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