PhosphoPlus^®RIP3 (Ser227) Antibody Duet#54105,Cell Signaling Technology (CST),54105

The receptor-interacting protein (RIP) family of serine-threonine kinases (RIP, RIP2, RIP3, and RIP4) are important regulators of cellular stress that trigger pro-survival and inflammatory responses through the activation of NF-κB, as well as pro-apoptotic pathways (1). In addition to the kinase domain, RIP contains a death domain responsible for interaction with the death domain receptor Fas and recruitment to TNF-R1 through interaction with TRADD (2,3). RIP-deficient cells show a failure in TNF-mediated NF-κB activation, making the cells more sensitive to apoptosis (4,5). RIP also interacts with TNF-receptor-associated factors (TRAFs) and can recruit IKKs to the TNF-R1 signaling complex via interaction with NEMO, leading to IκB phosphorylation and degradation (6,7). Overexpression of RIP induces both NF-κB activation and apoptosis (2,3). Caspase-8-dependent cleavage of the RIP death domain can trigger the apoptotic activity of RIP (8).Receptor-interacting protein 3 (RIP3) was originally found to interact with RIP and the TNF receptor complex to induce apoptosis and activation of NF-κB (9,10). Subsequently, it has been shown that the association between RIP and RIP3 is a key component of a signaling pathway that results in programmed necrosis (necroptosis), a necrotic-like cell death induced by TNF in the presence of caspase inhibitors (11-13). RIP3 is phosphorylated at Ser227 and targets the phosphorylation of mixed lineage kinase domain-like protein (MLKL), which is critical for necroptosis (14). In mice, RIP3 is phosphorylated at Thr231 and Ser232, leading to association with MLKL and necroptosis (15).Meylan, E. and Tschopp, J. (2005)Trends Biochem Sci30, 151-9.Hsu, H. et al. (1996)Immunity4, 387-96.Stanger, B.Z. et al. (1995)Cell81, 513-23.Ting, A.T. et al. (1996)EMBO J15, 6189-96.Kelliher, M.A. et al. (1998)Immunity8, 297-303.Devin, A. et al. (2000)Immunity12, 419-29.Zhang, S.Q. et al. (2000)Immunity12, 301-11.Lin, Y. et al. (1999)Genes Dev13, 2514-26.Yu, P.W. et al. (1999)Curr Biol9, 539-42.Sun, X. et al. (1999)J Biol Chem274, 16871-5.Zhang, D.W. et al. (2009)Science325, 332-6.He, S. et al. (2009)Cell137, 1100-11.Cho, Y.S. et al. (2009)Cell137, 1112-23.Sun, L. et al. (2012)Cell148, 213-27.Chen, W. et al. (2013)J Biol Chem288, 16247-61.