Fig. 1 | Nature Communications

Fig. 1

From: Conformational switching of the pseudokinase domain promotes human MLKL tetramerization and cell death by necroptosis

Fig. 1

Tetrameric hMLKL(2–471) is destabilized by ATP or PsK domain mutation to a monomeric intermediate. a Homology model of hMLKL21. The αC helix is highlighted green, the VAIK230 maroon, HGK331 blue, GFE351 orange, and the RIPK3 phosphorylation site, Thr357/Ser358, in purple. b, c Standardized continuous sedimentation coefficient [c(s20,w)] distribution of wild-type and E351K hMLKL(2–471) in the presence (red) and absence (blue) of 300 μM ATP at 1 mg/mL. Residuals for the best fit of the raw radial absorbance sedimentation velocity data to a continuous sedimentation coefficient [c(s)] distribution model are shown as an inset for corresponding protein at 1 mg/mL in the absence of ATP (top) and presence of 300 μM ATP (bottom). d, e Mass spectra of wild-type and E351K hMLKL(2–471) under native conditions. f Liposomes containing self-quenching dye 5(6)-carboxyfluorescein were added to 1 μM of wild-type hMLKL (dark blue), wild-type hMLKL + ATP (red), E351K hMLKL (orange), E351K hMLKL + ATP (light blue), and hMLKL(2-154) (green), and dye release was monitored by spectrophotometry at 485 nm over 30 min. Data represent mean ± SEM of three independent assays

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