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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

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MLKL, the Protein that Mediates Necroptosis, Also Regulates Endosomal Trafficking and Extracellular Vesicle

Seongmin Yoon1, Andrew Kovalenko1, Konstantin Bogdanov1

  • 1Department of Biomolecular Sciences, The Weizmann Institute of Science, 76100 Rehovot, Israel.

Immunity
|July 2, 2017
PubMed

Insights

Mixed lineage kinase domain-like (MLKL) protein controls endosomal transport and receptor degradation. RIPK3-activated MLKL also binds ESCRT proteins, facilitating its release in extracellular vesicles to self-restrict necroptosis.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Immunology

Background:

  • Mixed lineage kinase domain-like (MLKL) is a pseudokinase activated by RIPK3 phosphorylation, triggering necroptosis.
  • Necroptosis is a programmed cell death pathway involving membrane rupture and release of intracellular components.

Purpose of the Study:

  • To investigate the non-necroptotic functions of MLKL.
  • To elucidate the role of MLKL in endosomal transport and its regulation by RIPK3.

Main Methods:

  • Co-immunoprecipitation assays to identify MLKL interacting proteins.
  • Endocytosis and receptor degradation assays.
  • Confocal microscopy to visualize MLKL localization and vesicle formation.
  • Analysis of extracellular vesicle content.

Main Results:

  • MLKL constitutively associates with endosomes, controlling endocytosed protein transport, receptor/ligand degradation, and signaling modulation.
  • RIPK3 activation enhances MLKL association with endosomes, promoting binding to ESCRT proteins and flotillins.
  • MLKL, ESCRT, and flotillins are released from cells within extracellular vesicles upon RIPK3 activation.

Conclusions:

  • MLKL possesses a dual role: constitutive endosomal function and RIPK3-dependent regulation of endosomal trafficking.
  • Phosphorylated MLKL release via extracellular vesicles may serve as a negative feedback mechanism to limit necroptosis.
  • These findings reveal novel functions of MLKL beyond its canonical role in necroptosis.
Keywords:
EGFMLKLRIPK3TNFendosomesexosomesextracellular vesiclesinflammationintraluminal vesiclesmultivesicular bodiesnecroptosis

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