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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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Mixed lineage kinase domain-like protein MLKL causes necrotic membrane disruption upon phosphorylation by RIP3

Huayi Wang1, Liming Sun1, Lijing Su2

  • 1National Institute of Biological Sciences, No. 7 Science Park Road, Zhongguancun Life Science Park, Beijing 102206, China.

Molecular Cell
|April 8, 2014
PubMed

Insights

Necrotic cell death relies on RIP1 and RIP3 kinases. A new antibody detects phosphorylated MLKL, revealing its membrane-binding and cell-disrupting role in programmed necrosis.

Area of Science:

  • Cellular biology
  • Molecular mechanisms of cell death
  • Immunology

Background:

  • Programmed necrosis is initiated by tumor necrosis factor alpha (TNF-α) signaling.
  • This cell death pathway depends on receptor-interacting kinases RIP1 and RIP3.
  • The precise mechanism by which these kinases induce necrotic cell death remains unclear.

Purpose of the Study:

  • To investigate the role of mixed lineage kinase domain-like protein (MLKL) in TNF-α-induced programmed necrosis.
  • To develop a tool for detecting activated MLKL in cellular and clinical samples.
  • To elucidate the molecular events linking MLKL activation to membrane damage and cell death.

Main Methods:

  • Development of a monoclonal antibody specific for phosphorylated MLKL (pMLKL).
  • Analysis of pMLKL localization and oligomerization in dying cells.
  • Biochemical assays to determine the membrane-binding properties of pMLKL.
  • Examination of pMLKL in human liver biopsy samples from drug-induced liver injury cases.

Main Results:

  • A monoclonal antibody was generated that specifically recognizes phosphorylated MLKL (T357/S358).
  • Phosphorylated MLKL forms oligomers that bind to phosphatidylinositol lipids and cardiolipin.
  • This binding facilitates MLKL translocation from the cytosol to cellular membranes.
  • MLKL oligomers directly disrupt membrane integrity, leading to necrotic cell death.

Conclusions:

  • Phosphorylated MLKL is a key effector molecule in programmed necrosis.
  • MLKL's ability to bind membrane lipids and oligomerize is crucial for its cytotoxic function.
  • This pathway is implicated in human liver diseases, such as drug-induced liver injury.

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