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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

The pseudokinase MLKL mediates necroptosis via a molecular switch mechanism

James M Murphy1, Peter E Czabotar, Joanne M Hildebrand

  • 1The Walter and Eliza Hall Institute of Medical Research, Parkville, VIC 3052, Australia; Department of Medical Biology, University of Melbourne, Parkville, VIC 3050, Australia.

Immunity
|September 10, 2013
PubMed

Insights

Mixed lineage kinase domain-like (MLKL) is crucial for tumor necrosis factor-induced necroptosis. MLKL-deficient cells resist this cell death pathway, highlighting MLKL

Area of Science:

  • Cellular Biology
  • Molecular Biology
  • Immunology

Background:

  • Mixed lineage kinase domain-like (MLKL) is a key component of the necrosome complex.
  • The necrosome complex mediates tumor necrosis factor (TNF)-induced necroptosis, a form of programmed cell death.

Purpose of the Study:

  • To elucidate the specific role and molecular mechanism of MLKL in necroptosis.
  • To generate MLKL-deficient mice for functional studies and determine the MLKL structure.

Main Methods:

  • Generation and analysis of MLKL-deficient mice.
  • X-ray crystallography to solve the MLKL protein structure.
  • Site-directed mutagenesis to probe MLKL pseudoactive site function.

Main Results:

  • MLKL-deficient cells are resistant to TNF-induced necroptosis, confirming MLKL's essential role.
  • The crystal structure revealed MLKL's pseudokinase domain binds ATP but is catalytically inactive.
  • RIPK3-mediated phosphorylation induces MLKL's nonenzymatic function in necroptosis.
  • Mutations in the MLKL pseudoactive site lead to RIPK3-independent necroptosis.

Conclusions:

  • MLKL is indispensable for TNF-induced necroptosis signaling.
  • The pseudokinase domain's activity is regulated by RIPK3 phosphorylation, not intrinsic catalysis.
  • MLKL modification is critical for propagating the necroptosis pathway downstream of RIPK3.

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