The MLKL kinase-like domain dimerization is an indispensable step of mammalian MLKL activation in necroptosis

Related Experiment Video

Updated: Nov 1, 2025

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

11.0K

Yu Zhang1,2,3, Jia Liu1,3, Dandan Yu1,3

  • 1School of Life Science and Technology, ShanghaiTech University, 393 Middle Huaxia Road, Pudong, Shanghai, 201210, China.

Cell Death & Disease
|June 23, 2021
PubMed

Insights

Necroptosis execution involves mixed lineage kinase domain-like (MLKL) protein activation. This study reveals MLKL activation sequentially begins with kinase-like domain dimerization, followed by coiled-coil self-assembly for oligomerization and cell death.

Area of Science:

  • Cellular biology
  • Molecular biology
  • Biochemistry

Background:

  • Necroptosis is a regulated form of necrosis crucial in inflammation and immunity.
  • Mixed lineage kinase domain-like (MLKL) protein activation, triggered by RIP3-mediated phosphorylation, is essential for necroptosis execution.
  • MLKL activation involves a transition from monomeric to oligomeric forms, but the precise steps remain incompletely understood.

Purpose of the Study:

  • To elucidate the sequential molecular events governing MLKL activation during necroptosis.
  • To investigate the role of MLKL kinase-like domain dimerization and coiled-coil self-assembly in MLKL oligomerization and function.
  • To determine if MLKL kinase-like domain dimerization is conserved across mammalian species.

Main Methods:

  • X-ray crystallography to determine the structure of human MLKL.
  • Site-directed mutagenesis to disrupt identified MLKL interfaces and coiled-coil regions.
  • Functional assays to assess MLKL oligomerization and necroptosis in response to RIP3 activation.

Main Results:

  • The crystal structure of human MLKL revealed two inter-dimer interfaces within the kinase-like domain.
  • Mutations disrupting both inter-dimer interfaces abolished RIP3-induced MLKL oligomerization and necroptosis.
  • MLKL self-assembly via its internal coiled-coil region was necessary for oligomerization and necroptosis, but not for initial kinase-like domain dimerization.
  • Kinase-like domain dimerization was conserved in MLKL from other mammalian species.

Conclusions:

  • MLKL activation is a sequential process initiated by RIP3-induced kinase-like domain dimerization.
  • Subsequent self-assembly of the internal coiled-coil region leads to the formation of functional MLKL oligomers.
  • This sequential activation mechanism, starting with kinase-like domain dimerization, is conserved in mammals, highlighting its fundamental role in necroptosis.

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