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

Related Experiment Video

Updated: Feb 8, 2026

Characterization of MLKL-mediated Plasma Membrane Rupture in Necroptosis
08:55

Characterization of MLKL-mediated Plasma Membrane Rupture in Necroptosis

Published on: August 7, 2018

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Emma J Petrie1,2, Jarrod J Sandow3,4, Annette V Jacobsen3,4

  • 1The Walter & Eliza Hall Institute of Medical Research, Parkville, VIC, Australia. petrie.e@wehi.edu.au.

Nature Communications
|June 23, 2018
PubMed

Insights

This study reveals how the MLKL protein activates necroptotic cell death. It identifies a key conformational change in MLKL, crucial for initiating this cell death pathway and differing between human and mouse.

Area of Science:

  • Cellular Biology
  • Molecular Mechanisms of Cell Death

Background:

  • Necroptotic cell death is a regulated form of cell death.
  • MLKL (mixed lineage kinase domain-like) is the terminal effector of necroptosis.
  • The precise activation mechanism of MLKL by RIPK3 remains unclear.

Purpose of the Study:

  • To elucidate the structural and mechanistic basis of MLKL activation in necroptosis.
  • To understand the role of MLKL pseudokinase domain phosphorylation by RIPK3.
  • To compare MLKL activation mechanisms between human and mouse.

Main Methods:

  • Analysis of MLKL conformation in human cells.
  • Investigating the role of RIPK3 phosphorylation sites in human MLKL.
  • Comparative studies of human and mouse MLKL activation.

Main Results:

  • Identified a basal monomeric MLKL conformer in human cells.
  • Demonstrated that MLKL activation involves a conformational switch within the pseudokinase domain.
  • Showed that RIPK3 substrate site mutations in human MLKL abolish necroptosis, unlike in mouse MLKL.
  • Revealed species-specific differences in MLKL activation mechanisms.

Conclusions:

  • MLKL acts as a molecular switch, regulated by its pseudokinase domain.
  • The N-terminal 4HB domain disengages upon activation, enabling tetramer formation and membrane permeabilization.
  • Human and mouse MLKL activation pathways, while both involving pseudokinase domain switching, exhibit critical mechanistic differences.

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