Mixed lineage kinase domain-like protein MLKL causes necrotic membrane disruption upon phosphorylation by RIP3.
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.
National Institute of Biological Sciences, No. 7 Science Park Road, Zhongguancun Life Science Park, Beijing 102206, China.
Huayi
H
Wang
National Institute of Biological Sciences, No. 7 Science Park Road, Zhongguancun Life Science Park, Beijing 102206, China.
Liming
L
Sun
Departments of Biophysics, Biochemistry and Pharmacology, University of Texas Southwestern Medical Center, 6000 Harry Hines Boulevard, Dallas, TX 75390, USA.
Lijing
L
Su
Departments of Biophysics, Biochemistry and Pharmacology, University of Texas Southwestern Medical Center, 6000 Harry Hines Boulevard, Dallas, TX 75390, USA.
Josep
J
Rizo
National Institute of Biological Sciences, No. 7 Science Park Road, Zhongguancun Life Science Park, Beijing 102206, China.
Lei
L
Liu
Research Center for Biological Therapy, The Institute of Translational Hepatology, Beijing 302 Hospital, Beijing 100039, China.
Li-Feng
LF
Wang
Research Center for Biological Therapy, The Institute of Translational Hepatology, Beijing 302 Hospital, Beijing 100039, China.
Fu-Sheng
FS
Wang
National Institute of Biological Sciences, No. 7 Science Park Road, Zhongguancun Life Science Park, Beijing 102206, China. Electronic address: wangxiaodong@nibs.ac.cn.
Xiaodong
X
Wang
D000911:Antibodies, Monoclonal / Q000276:immunology; D001665:Binding Sites; D002308:Cardiolipins / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology* / Q000473:pathology; D056486:Chemical and Drug Induced Liver Injury / Q000201:enzymology / Q000473:pathology; D019073:HT29 Cells; D006367:HeLa Cells; D006801:Humans; D007425:Intracellular Membranes / Q000201:enzymology / Q000473:pathology; D008099:Liver / Q000201:enzymology / Q000473:pathology; D008563:Membrane Lipids / Q000378:metabolism; D009336:Necrosis; D010766:Phosphorylation; D011487:Protein Conformation; D011494:Protein Kinases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D021381:Protein Transport; D034622:RNA Interference; D053422:Receptor-Interacting Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D013379:Substrate Specificity; D013997:Time Factors; D014162:Transfection
D000911:Antibodies, Monoclonal; D002308:Cardiolipins; D008563:Membrane Lipids; C570693:MLKL protein, human; D011494:Protein Kinases; C119705:RIPK3 protein, human; D053422:Receptor-Interacting Protein Serine-Threonine Kinases