Abstract
TWEAK, like many other ligands of the TNF family, occurs naturally in two forms, as a type II transmembrane protein and as soluble ligand released from the latter by proteases of the furin family. Both TWEAK variants interact with high affinity with Fn14, an unusual small member of the TNF receptor family. TWEAK and Fn14 activate a variety of intracellular signaling pathways but regulation of TNF-induced cell death and stimulation of the classical and alternative NFκB pathway are certainly the best understood ones. Intriguingly, soluble and membrane TWEAK significantly differ in their ability to trigger these responses. While activation of the alternative NFκB pathway and enhancement of TNF-induced cell death are efficiently induced by both forms of TWEAK, membrane TWEAK has a much higher capacity than soluble TWEAK to stimulate the classical NFκB pathway. Importantly, soluble TWEAK gains a membrane TWEAK-like Fn14 stimulating activity upon oligomerization or artificial anchoring to the cell surface. On the example of NFκB signaling and enhancement of TNF-induced cell death, we summarize here protocols that allow the identification of signaling pathways/cellular responses that preferentially respond to membrane TWEAK. These protocols base either on the side-by-side analysis of soluble TWEAK and oligomerized or cell surface-anchorable TWEAK variants or on the use of transfectants expressing soluble and membrane TWEAK.
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References
Burkly LC, Michaelson JS, Zheng TS (2011) TWEAK/Fn14 pathway: an immunological switch for shaping tissue responses. Immunol Rev 244:99–114
Winkles JA (2008) The TWEAK-Fn14 cytokine-receptor axis: discovery, biology and therapeutic targeting. Nat Rev Drug Discov 7:411–425
Grell M, Douni E, Wajant H, Löhden M, Clauss M, Maxeiner B et al (1995) The transmembrane form of tumor necrosis factor is the prime activating ligand of the 80 kDa tumor necrosis factor receptor. Cell 83:793–802
Schneider P, Holler N, Bodmer JL, Hahne M, Frei K, Fontana A et al (1998) Conversion of membrane-bound Fas(CD95) ligand to its soluble form is associated with downregulation of its proapoptotic activity and loss of liver toxicity. J Exp Med 187:1205–1213
Wajant H, Moosmayer D, Wüest T, Bartke T, Gerlach E, Schönherr U et al (2001) Differential activation of TRAIL-R1 and -2 by soluble and membrane TRAIL allows selective surface antigen-directed activation of TRAIL-R2 by a soluble TRAIL derivative. Oncogene 20:4101–4106
Müller N, Wyzgol A, Münkel S, Pfizenmaier K, Wajant H (2008) Activity of soluble OX40 ligand is enhanced by oligomerization and cell surface immobilization. FEBS J 275:2296–22304
Wyzgol A, Müller N, Fick A, Munkel S, Grigoleit GU, Pfizenmaier K et al (2009) Trimer stabilization, oligomerization, and antibody-mediated cell surface immobilization improve the activity of soluble trimers of CD27L, CD40L, 41BBL, and glucocorticoid-induced TNF receptor ligand. J Immunol 183:1851–1861
Bossen C, Cachero TG, Tardivel A, Ingold K, Willen L, Dobles M et al (2008) TACI, unlike BAFF-R, is solely activated by oligomeric BAFF and APRIL to support survival of activated B cells and plasmablasts. Blood 111:1004–1012
Schneider P, Schwenzer R, Haas E, Mühlenbeck F, Schubert G, Scheurich P et al (1999) TWEAK can induce cell death via endogenous TNF and TNF receptor 1. Eur J Immunol 29:1785–1792
Wicovsky A, Salzmann S, Roos C, Ehrenschwender M, Rosenthal T, Siegmund D et al (2009) TNF-like weak inducer of apoptosis inhibits proinflammatory TNF receptor-1 signaling. Cell Death Differ 16:1445–1459
Roos C, Wicovsky A, Müller N, Salzmann S, Rosenthal T, Kalthoff H et al (2010) Soluble and transmembrane TNF-like weak inducer of apoptosis differentially activate the classical and noncanonical NF-kappa B pathway. J Immunol 185:1593–1605
Fick A, Lang I, Schäfer V, Seher A, Trebing J, Weisenberger D et al (2012) Studies of binding of tumor necrosis factor (TNF)-like weak inducer of apoptosis (TWEAK) to fibroblast growth factor inducible 14 (Fn14). J Biol Chem 287:484–495
Bover LC, Cardó-Vila M, Kuniyasu A, Sun J, Rangel R, Takeya M et al (2007) A previously unrecognized protein-protein interaction between TWEAK and CD163: potential biological implications. J Immunol 178:8183–8194
Fick A, Wyzgol A, Wajant H (2012) Production, purification, and characterization of scFv TNF ligand fusion proteins. Methods Mol Biol 907:597–609
Brown SA, Ghosh A, Winkles JA (2010) Full-length, membrane-anchored TWEAK can function as a juxtacrine signaling molecule and activate the NF-kappaB pathway. J Biol Chem 285:17432–17441
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Trebing, J., Arana, J.A.C., Salzmann, S., Wajant, H. (2014). Analyzing the Signaling Capabilities of Soluble and Membrane TWEAK. In: Bayry, J. (eds) The TNF Superfamily. Methods in Molecular Biology, vol 1155. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0669-7_4
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DOI: https://doi.org/10.1007/978-1-4939-0669-7_4
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