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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Priem, D., Dondelinger, Y., Bertrand, M.J.M. (2018). Monitoring RIPK1 Phosphorylation in the TNFR1 Signaling Complex. In: Ting, A. (eds) Programmed Necrosis. Methods in Molecular Biology, vol 1857. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8754-2_17
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DOI: https://doi.org/10.1007/978-1-4939-8754-2_17
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