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

Mol Cell. 2014 Apr 10;54(1):133-146. doi: 10.1016/j.molcel.2014.03.003. Epub 2014 Apr 3.


Programmed necrotic cell death induced by the tumor necrosis factor alpha (TNF-α) family of cytokines is dependent on a kinase cascade consisting of receptor-interacting kinases RIP1 and RIP3. How these kinase activities cause cells to die by necrosis is not known. The mixed lineage kinase domain-like protein MLKL is a functional RIP3 substrate that binds to RIP3 through its kinase-like domain but lacks kinase activity of its own. RIP3 phosphorylates MLKL at the T357 and S358 sites. Reported here is the development of a monoclonal antibody that specifically recognizes phosphorylated MLKL in cells dying of this pathway and in human liver biopsy samples from patients suffering from drug-induced liver injury. The phosphorylated MLKL forms an oligomer that binds to phosphatidylinositol lipids and cardiolipin. This property allows MLKL to move from the cytosol to the plasma and intracellular membranes, where it directly disrupts membrane integrity, resulting in necrotic death.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / immunology
  • Binding Sites
  • Cardiolipins / metabolism
  • Cell Membrane / enzymology*
  • Cell Membrane / pathology
  • Chemical and Drug Induced Liver Injury / enzymology
  • Chemical and Drug Induced Liver Injury / pathology
  • HT29 Cells
  • HeLa Cells
  • Humans
  • Intracellular Membranes / enzymology
  • Intracellular Membranes / pathology
  • Liver / enzymology
  • Liver / pathology
  • Membrane Lipids / metabolism
  • Necrosis
  • Phosphorylation
  • Protein Conformation
  • Protein Kinases / genetics
  • Protein Kinases / immunology
  • Protein Kinases / metabolism*
  • Protein Transport
  • RNA Interference
  • Receptor-Interacting Protein Serine-Threonine Kinases / genetics
  • Receptor-Interacting Protein Serine-Threonine Kinases / metabolism*
  • Signal Transduction
  • Substrate Specificity
  • Time Factors
  • Transfection


  • Antibodies, Monoclonal
  • Cardiolipins
  • Membrane Lipids
  • MLKL protein, human
  • Protein Kinases
  • RIPK3 protein, human
  • Receptor-Interacting Protein Serine-Threonine Kinases