Fig. 6: RIPK1 is SUMOyalted at K550 and SUMOylation of RIPK1 promotes its activation.
From: SENP1 prevents steatohepatitis by suppressing RIPK1-driven apoptosis and inflammation
a Multiple sequence alignment of RIPK1 proteins from different species. K550 of murine RIPK1 (K565 in human RIPK1) locates within SUMO modification consensus motif (ψ-K-×E/D), which is highly conserved among different species, but not that of K132 of RIPK1. ψ: hydrophobic amino acid; K: the target lysine; ×: any amino acid; E or D is an acidic residue. b MEFs were stimulated by Flag-TNFα (100 ng/ml) for indicated time. SUMOylated RIPK1 was isolated by tandem immunoprecipitation of TNF-RSC, and then analyzed by immunoblotting with anti-RIPK1 antibody. MEFs were stimulated by Flag-TNFα (100 ng/ml) for indicated time. TNF-RSC was immunoprecipitated using anti-Flag resin. The immune complexes were analyzed by immunoblotting using anti-RIPK1 antibody and anti-p-S166 RIPK1 antibody (c), and anti-A20 antibody and other antibodies as indicated (d). MEFs were treated with TNFα (10 ng/ml) in the presence or absence of Nec-1s (10 μM). Cell death was measured as a function of time by SytoxGreen positivity assay (e). The level of p-S166 RIPK1 and CC3 were determined by immunoblotting (f). n = 3 biologically independent experiments (b–f). Data are represented as mean ± s.d. (e). Source data are provided in Source Data file.
