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Monitoring RIPK1 phosphorylation in the TNFR1 signaling complex

Dario Priem (UGent) , Yves Dondelinger (UGent) and Mathieu Bertrand (UGent)
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Abstract
Receptor-interacting protein kinase 1 (RIPK1) is a component of the TNFR1 signaling complex (also known as complex I or TNFR-SC), where its ubiquitylation by cIAP1/2 and LUBAC serves to initiate prosurvival and proinflammatory responses through activation of the MAPK and NF-κB pathways. IKKα/β-mediated phosphorylation of RIPK1 in complex I was shown to maintain RIPK1 in a prosurvival modus. Consequently, conditions affecting proper IKKα/β activation perturb IKKα/β-phosphorylation of RIPK1 and switch the TNF response toward RIPK1 kinase-dependent cell death. Methods to study the posttranslational modifications of RIPK1 in complex I are therefore of great value. Here, we describe a detailed protocol to isolate complex I-associated RIPK1 from cells and provide different tools to study the phosphorylation status of RIPK1 in TNFR1 complex I.

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MLA
Priem, Dario, et al. “Monitoring RIPK1 Phosphorylation in the TNFR1 Signaling Complex.” Programmed Necrosis : Methods and Protocols, edited by Adrian T Ting, vol. 1857, Springer Humana Press, 2018, pp. 171–79, doi:10.1007/978-1-4939-8754-2_17.
APA
Priem, D., Dondelinger, Y., & Bertrand, M. (2018). Monitoring RIPK1 phosphorylation in the TNFR1 signaling complex. In A. T. Ting (Ed.), Programmed necrosis : methods and protocols (Vol. 1857, pp. 171–179). https://doi.org/10.1007/978-1-4939-8754-2_17
Chicago author-date
Priem, Dario, Yves Dondelinger, and Mathieu Bertrand. 2018. “Monitoring RIPK1 Phosphorylation in the TNFR1 Signaling Complex.” In Programmed Necrosis : Methods and Protocols, edited by Adrian T Ting, 1857:171–79. New York, NY, USA: Springer Humana Press. https://doi.org/10.1007/978-1-4939-8754-2_17.
Chicago author-date (all authors)
Priem, Dario, Yves Dondelinger, and Mathieu Bertrand. 2018. “Monitoring RIPK1 Phosphorylation in the TNFR1 Signaling Complex.” In Programmed Necrosis : Methods and Protocols, ed by. Adrian T Ting, 1857:171–179. New York, NY, USA: Springer Humana Press. doi:10.1007/978-1-4939-8754-2_17.
Vancouver
1.
Priem D, Dondelinger Y, Bertrand M. Monitoring RIPK1 phosphorylation in the TNFR1 signaling complex. In: Ting AT, editor. Programmed necrosis : methods and protocols. New York, NY, USA: Springer Humana Press; 2018. p. 171–9.
IEEE
[1]
D. Priem, Y. Dondelinger, and M. Bertrand, “Monitoring RIPK1 phosphorylation in the TNFR1 signaling complex,” in Programmed necrosis : methods and protocols, vol. 1857, A. T. Ting, Ed. New York, NY, USA: Springer Humana Press, 2018, pp. 171–179.
@incollection{8582671,
  abstract     = {{Receptor-interacting protein kinase 1 (RIPK1) is a component of the TNFR1 signaling complex (also known as complex I or TNFR-SC), where its ubiquitylation by cIAP1/2 and LUBAC serves to initiate prosurvival and proinflammatory responses through activation of the MAPK and NF-κB pathways. IKKα/β-mediated phosphorylation of RIPK1 in complex I was shown to maintain RIPK1 in a prosurvival modus. Consequently, conditions affecting proper IKKα/β activation perturb IKKα/β-phosphorylation of RIPK1 and switch the TNF response toward RIPK1 kinase-dependent cell death. Methods to study the posttranslational modifications of RIPK1 in complex I are therefore of great value. Here, we describe a detailed protocol to isolate complex I-associated RIPK1 from cells and provide different tools to study the phosphorylation status of RIPK1 in TNFR1 complex I.}},
  author       = {{Priem, Dario and Dondelinger, Yves and Bertrand, Mathieu}},
  booktitle    = {{Programmed necrosis : methods and protocols}},
  editor       = {{Ting, Adrian T}},
  isbn         = {{9781493987535}},
  issn         = {{1064-3745}},
  language     = {{eng}},
  pages        = {{171--179}},
  publisher    = {{Springer Humana Press}},
  series       = {{Methods in Molecular Biology}},
  title        = {{Monitoring RIPK1 phosphorylation in the TNFR1 signaling complex}},
  url          = {{http://dx.doi.org/10.1007/978-1-4939-8754-2_17}},
  volume       = {{1857}},
  year         = {{2018}},
}

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