Affinage

MLKL

Mixed lineage kinase domain-like protein · UniProt Q8NB16

Length
471 aa
Mass
54.5 kDa
Annotated
2026-04-28
100 papers in source corpus 37 papers cited in narrative 37 extracted findings

Mechanistic narrative

Synthesis pass · prose summary of the discoveries below

MLKL is a pseudokinase that serves as the obligate terminal effector of necroptosis and additionally participates in endosomal trafficking, autophagy, phagocytosis, and innate immune signaling independently of cell death. In the necroptosis pathway, RIPK3 phosphorylates MLKL at T357/S358 (human), triggering pseudokinase domain dimerization, brace helix–nucleated tetramerization, release of the N-terminal four-helix bundle (4HB) domain, and sequential engagement of phosphatidylinositol lipids—preferentially PI(4,5)P2—enabling Golgi–microtubule–actin-dependent translocation to the plasma membrane where MLKL oligomers permeabilize the bilayer, induce K⁺ efflux, activate the NLRP3 inflammasome, and trigger release of DAMPs and mitochondrial DNA that engages the cGAS–STING–IFN-β axis (PMID:24703947, PMID:37884510, PMID:26853145, PMID:32561730, PMID:28096356, PMID:40614706). MLKL activity is restrained by multi-mono-ubiquitylation and K63-linked ubiquitin chains (via ITCH and Skp2), ESCRT-III–mediated membrane shedding, and autoinhibition through its brace region (PMID:34698396, PMID:34999730, PMID:28388412, PMID:32561755). Beyond necroptosis, MLKL constitutively associates with endosomes to regulate receptor degradation and extracellular vesicle release, is phosphorylated by CaMKII to promote autophagosome–lysosome fusion during starvation, facilitates macrophage phagocytosis via STAT1-dependent induction, and undergoes S441 phosphorylation in Schwann cells to drive myelin breakdown after nerve injury (PMID:28666573, PMID:34282994, PMID:35689613, PMID:30344099).

Mechanistic history

Synthesis pass · year-by-year structured walk · 16 steps
  1. 2013 High

    Resolving whether MLKL is a functional kinase or a pseudokinase established the fundamental paradigm: MLKL binds ATP but lacks catalytic activity, functioning instead as a conformational switch activated by RIPK3 phosphorylation, with MLKL-deficient mice resistant to TNF-induced necroptosis.

    Evidence Crystal structure of full-length mouse MLKL, active-site mutagenesis, MLKL knockout mice

    PMID:24012422

    Open questions at the time
    • Structural basis of full-length human MLKL not yet resolved
    • Mechanism of membrane permeabilization unknown at this stage
  2. 2013 High

    Epistasis experiments placed MLKL as the terminal effector downstream of RIPK3 across multiple death receptor and TLR-initiated necroptosis pathways, including RIP1-independent branches.

    Evidence Genetic epistasis with RIP1-KO, RIP3-KO, MLKL-KD in TLR3/TLR4-TRIF signaling in fibroblasts

    PMID:24019532

    Open questions at the time
    • Mechanism by which MLKL executes membrane disruption not defined
  3. 2014 High

    Identification of T357/S358 as the RIPK3 phosphorylation sites on MLKL, and demonstration that phosphorylation triggers oligomerization, phospholipid binding, and membrane translocation, established the core biochemical mechanism of necroptosis execution.

    Evidence In vitro kinase assay, phospho-specific antibodies, lipid-binding assays, cell fractionation, loss-of-function studies

    PMID:24577084 PMID:24703947

    Open questions at the time
    • Oligomeric stoichiometry undefined
    • Mechanism of lipid selectivity unknown
    • Drp1/mitochondrial fission confirmed dispensable but other organelle contributions unclear
  4. 2016 High

    Defining PI(4,5)P2 as MLKL's preferred lipid target and revealing a sequential 'rolling over' mechanism of lipid engagement by the 4HB domain explained how MLKL transitions from cytosolic to membrane-embedded killer, and cross-species analysis showed evolutionary divergence in 4HB killing competence.

    Evidence In vitro lipid-binding assays with mutagenesis, liposome permeabilization reconstitution across species, forced dimerization constructs

    PMID:26853145 PMID:26868910

    Open questions at the time
    • Why human 4HB requires dimerization while mouse does not is mechanistically unexplained
    • In vivo lipid requirement not tested
  5. 2016 High

    Discovery of RIPK3-independent MLKL activation in experimental hepatitis, driven by IFN-γ/STAT1-mediated transcriptional upregulation, established that MLKL has pathological roles beyond canonical RIPK3-dependent necroptosis.

    Evidence MLKL-KO vs RIPK3-KO mice in ConA hepatitis model, STAT1 pathway analysis, human biopsy validation

    PMID:27756058

    Open questions at the time
    • Kinase responsible for MLKL phosphorylation in RIPK3-independent hepatitis not identified
    • Generalizability to other tissues unknown
  6. 2017 High

    Demonstration that active MLKL triggers K⁺ efflux-dependent NLRP3 inflammasome assembly and caspase-1-mediated IL-1β processing before cell lysis linked necroptosis execution to inflammasome-driven inflammation.

    Evidence Chemogenetic MLKL activation, NLRP3/caspase-1/GSDMD KO cells, K⁺ measurement, ASC speck imaging

    PMID:28096356 PMID:28130493

    Open questions at the time
    • Whether MLKL-dependent NLRP3 activation occurs physiologically in vivo independent of other DAMPs not established
  7. 2017 High

    Identification of ESCRT-III as a membrane repair/shedding mechanism that antagonizes MLKL pore formation, and discovery that MLKL constitutively localizes to endosomes independently of RIPK3, revealed that MLKL has non-death functions in endosomal trafficking and extracellular vesicle biogenesis.

    Evidence ESCRT-III component KO with live imaging of PS exposure/Ca²⁺ flux; MLKL-KO cells with endosome fractionation, vesicle isolation, ESCRT/flotillin Co-IP

    PMID:28388412 PMID:28666573

    Open questions at the time
    • Structural basis of MLKL-ESCRT interaction unknown
    • Endosomal function of MLKL in vivo not independently validated
  8. 2018 High

    Biochemical dissection of human MLKL showed that RIPK3 phosphorylation disengages the 4HB domain from the pseudokinase domain αC helix, enabling tetramer formation—a species-specific requirement absent in mouse MLKL—while TAM kinases were identified as additional MLKL phosphorylators at Y376 that promote oligomerization.

    Evidence Native PAGE, SEC-MALS, mutagenesis for human vs mouse constructs; TAM kinase KO and inhibition with phospho-site mutagenesis and in vivo SIRS model

    PMID:29930286 PMID:31230815

    Open questions at the time
    • Relative contribution of TAM vs RIPK3 phosphorylation in physiological necroptosis unclear
    • Human MLKL tetramer structure not yet solved at this stage
  9. 2018 High

    Discovery that MLKL undergoes distinct S441 phosphorylation in Schwann cells after nerve injury to target myelin membranes, and that MLKL activates PAD4-dependent NET formation in neutrophils, broadened MLKL's functional repertoire to tissue repair and innate immunity.

    Evidence Schwann cell-specific Mlkl KO in sciatic nerve injury model; MLKL-KO and PAD4-KO neutrophils with NET quantification

    PMID:30181240 PMID:30344099

    Open questions at the time
    • Kinase responsible for S441 phosphorylation in Schwann cells not identified
    • Whether MLKL-driven NETosis is mechanistically distinct from MLKL-driven necroptosis at the membrane level is unclear
  10. 2020 High

    Live imaging of endogenous MLKL revealed that phosphorylated MLKL co-traffics with tight junction proteins via Golgi–microtubule–actin pathways to plasma membrane hotspots, establishing intracellular trafficking as a regulated necroptosis checkpoint; a brace-region mutation (D139V) was shown to cause constitutive RIPK3-independent killing, identifying the brace as a critical autoinhibitory element.

    Evidence CRISPR-tagged endogenous MLKL with super-resolution imaging and pharmacological trafficking disruption; ENU mutagenesis screen with RIPK3-KO epistasis in vivo

    PMID:32561730 PMID:32561755

    Open questions at the time
    • Molecular basis of MLKL sorting into Golgi-dependent trafficking route unknown
    • How tight junction association relates to membrane permeabilization not resolved
  11. 2020 High

    Hsp70 was identified as a direct MLKL-binding partner that promotes amyloid-like polymerization of the MLKL N-terminal domain, providing a chaperone-dependent mechanism for MLKL higher-order assembly; MLKL was also shown to function in a RIPK3-independent, autophagy-inhibiting role in hepatocytes exposed to lipotoxic stress.

    Evidence Recombinant Hsp70-MLKL reconstitution, Cys mutagenesis, NBC1 inhibitor; Mlkl-KO vs Rip3-KO mice on Western diet with mRFP-GFP-LC3 autophagy reporter

    PMID:32156734 PMID:32220583

    Open questions at the time
    • Physiological relevance of amyloid-like MLKL polymers vs canonical oligomers in cell death not distinguished
    • Mechanism linking MLKL to autophagy blockade at the molecular level unclear
  12. 2021 High

    Structural studies resolved MLKL pseudokinase domain dimerization interfaces and showed conformational interconversion upon RIPK3 disengagement, while ubiquitylation was identified as a major post-translational restraint on MLKL: multi-mono-ubiquitylation targets membrane-associated MLKL for proteasomal/lysosomal turnover, and removal of ubiquitin licenses spontaneous MLKL activation.

    Evidence Crystal structures with Monobody tools and interface mutagenesis; mass spectrometry of ubiquitylation sites, MLKL-DUB fusion constructs, proteasome/lysosome inhibitors

    PMID:33850121 PMID:34158471 PMID:34698396

    Open questions at the time
    • Complete atomic structure of full-length activated MLKL oligomer not available
    • Identity of E3 ligase(s) for multi-mono-ubiquitylation not determined at this stage
  13. 2021 High

    CaMKII was identified as a RIPK3-independent kinase that phosphorylates MLKL during starvation to promote autophagosome–lysosome fusion, and MLKL was shown to support macrophage phagocytosis via STAT1-dependent expression and translocation to phagosomes/lysosomes, solidifying MLKL's non-necroptotic roles.

    Evidence CaMKII KO/inhibition with autophagy reporters in RIPK3-independent setting; bone marrow chimeras with Mlkl-KO defining myeloid-specific phagocytic requirement

    PMID:34282994 PMID:35689613

    Open questions at the time
    • CaMKII phosphorylation sites on MLKL not mapped
    • Whether MLKL's phagocytic function requires oligomerization or membrane binding is unknown
  14. 2022 High

    K63-linked ubiquitylation of the MLKL HeLo domain by E3 ligase ITCH was shown to divert phosphorylated MLKL from plasma membrane to endosomes, channeling intracellular bacteria to lysosomes for destruction, revealing ubiquitin chain topology as a trafficking determinant.

    Evidence ITCH Co-IP/pulldown, K63-specific ubiquitylation mutants, bacterial infection assays, extracellular vesicle isolation

    PMID:34999730

    Open questions at the time
    • Whether ITCH and Skp2 act on the same or distinct MLKL pools is unclear
    • Structural basis of ITCH–MLKL WW domain interaction not resolved
  15. 2023 High

    The full structural mechanism of human MLKL activation was resolved: RIPK3 phosphorylation drives pseudokinase domain dimerization that nucleates an elongated homotetramer via an ~80 Å brace helix coiled coil, enabling 4HB domain release and membrane permeabilization; MLKL polymers were additionally shown to target lysosomal membranes, causing cathepsin B-dependent cell death.

    Evidence Crystal structure and negative-stain EM of MLKL tetramer with mutagenesis; MLKL NTD expression with lysosomal imaging and cathepsin B inhibition

    PMID:37884510 PMID:37996483

    Open questions at the time
    • Cryo-EM structure of membrane-inserted MLKL pore not available
    • Relative contribution of plasma membrane vs lysosomal membrane disruption to necroptotic death not quantified
  16. 2025 High

    Phosphorylated MLKL was shown to translocate to mitochondria and induce microtubule-dependent mitochondrial DNA release into the cytosol, activating cGAS–STING and IFN-β, connecting MLKL's membrane-permeabilizing activity to innate immune DNA sensing.

    Evidence Subcellular fractionation, live imaging, mtDNA quantification, cGAS/STING-KO cells, in vivo IBD model

    PMID:40614706

    Open questions at the time
    • Whether MLKL forms the same oligomeric species on mitochondrial membranes as on plasma membrane is unknown
    • Relative in vivo contribution of MLKL-mtDNA-cGAS vs DAMP-mediated inflammation not dissected

Open questions

Synthesis pass · forward-looking unresolved questions
  • Major open questions include: the atomic structure of the membrane-inserted MLKL pore/channel, the identity of the kinase(s) responsible for RIPK3-independent MLKL phosphorylation in hepatitis and Schwann cells (S441), the precise CaMKII phosphorylation sites on MLKL, and how ubiquitin chain topology (mono vs K63) mechanistically determines MLKL trafficking fate between plasma membrane and endosomes.
  • No cryo-EM or cryo-ET structure of membrane-embedded MLKL pore
  • Kinase for S441 phosphorylation in Schwann cells unidentified
  • CaMKII target residues on MLKL unmapped
  • Decision logic between ITCH K63-ubiquitin-mediated endosomal routing and multi-mono-ubiquitin-mediated degradation not resolved

Mechanism profile

Synthesis pass · controlled-vocabulary classification · explore literature graph →
Molecular activity
GO:0005198 structural molecule activity 3 GO:0008289 lipid binding 3
Localization
GO:0005886 plasma membrane 6 GO:0005768 endosome 3 GO:0005829 cytosol 3 GO:0005764 lysosome 2 GO:0005634 nucleus 1 GO:0005739 mitochondrion 1 GO:0005794 Golgi apparatus 1
Pathway
R-HSA-5357801 Programmed Cell Death 7 R-HSA-168256 Immune System 6 R-HSA-162582 Signal Transduction 4 R-HSA-5653656 Vesicle-mediated transport 4 R-HSA-9612973 Autophagy 2
Partners
AXLCAMK2HSP70ITCHMERTKRIPK3SKP2TYRO3
Complex memberships
necrosome (RIPK3-MLKL)

Evidence

Reading pass · 37 per-paper findings extracted from the source corpus
Year Finding Method Journal Conf PMIDs
2014 MLKL is a direct substrate of RIP3 kinase; RIP3 phosphorylates MLKL at T357 and S358, which triggers MLKL oligomerization, binding to phosphatidylinositol lipids and cardiolipin, translocation from cytosol to plasma and intracellular membranes, and direct membrane disruption causing necrotic cell death. In vitro kinase assay, phospho-specific monoclonal antibody, Co-IP, lipid-binding assay, cell fractionation, loss-of-function Molecular cell High 24703947
2013 MLKL is a pseudokinase that binds ATP but is catalytically inactive; its pseudokinase domain acts as a molecular switch that is activated by RIPK3-mediated phosphorylation, and structure-guided mutation of the pseudoactive site causes constitutive, RIPK3-independent necroptosis. MLKL-deficient mice are resistant to TNF-induced necroptosis. Crystal structure of MLKL, active-site mutagenesis, MLKL knockout mice, cell death assays Immunity High 24012422
2016 MLKL executes necroptosis through sequential engagement of distinct phosphatidylinositol-binding sites: the N-terminal helix bundle (NB) first binds PIP lipids with low affinity to target the plasma membrane, then undergoes a 'rolling over' mechanism to expose higher-affinity PIP-binding sites; PI(4,5)P2 is the preferred binding partner. In vitro lipid-binding assays, structural analysis, mutagenesis, cell death assays Molecular cell High 26853145
2013 MLKL acts downstream of RIPK3 in TLR3/TLR4-TRIF-induced programmed necrosis, functioning independently of RIP1 or its kinase activity in fibroblasts, placing MLKL as the terminal effector in multiple necroptotic signaling branches. Genetic epistasis (RIP1 KO, RIP3 KO, MLKL knockdown), small-molecule RIP3 inhibitors, cell death assays The Journal of biological chemistry High 24019532
2017 ESCRT-III machinery acts downstream of MLKL activation to shed plasma membrane 'bubbles' with exposed phosphatidylserine; MLKL-dependent Ca2+ influx and PS exposure precede membrane rupture. ESCRT-III controls the kinetics of membrane integrity loss during necroptosis. Live-cell imaging, fractionation, genetic KO of ESCRT-III components, flow cytometry for PS exposure and Ca2+ flux Cell High 28388412
2017 MLKL also associates with endosomes constitutively (independent of RIPK3) and in enhanced form upon RIPK3 activation, controlling endocytosed protein transport, receptor/ligand degradation, and extracellular vesicle generation; phosphorylated MLKL binds ESCRT proteins and flotillins on endosomes and is released within extracellular vesicles. Co-IP, subcellular fractionation, vesicle isolation, live imaging, MLKL KO cells Immunity High 28666573
2020 Phosphorylated MLKL assembles into higher-order species on cytoplasmic necrosomes, then co-traffics with tight junction proteins to the cell periphery via Golgi-microtubule-actin-dependent mechanisms, accumulating as hotspots at the plasma membrane; this trafficking and accumulation are crucial necroptosis checkpoints. Single-cell live imaging of endogenous MLKL (CRISPR-tagged), super-resolution microscopy, pharmacological disruption of Golgi/microtubule/actin Nature communications High 32561730
2018 RIPK3-mediated phosphorylation of human MLKL induces a conformational switch in the pseudokinase domain causing disengagement of the 4HB domain from the pseudokinase domain αC helix and pseudocatalytic loop, enabling formation of a necroptosis-inducing tetramer; substitution of RIPK3 phosphorylation sites completely abrogates human (but not mouse) necroptotic signaling. Native PAGE, SEC-MALS, mutagenesis, cell death assays, comparison of human vs. mouse MLKL constructs Nature communications High 29930286
2019 TAM receptor tyrosine kinases (Tyro3, Axl, Mer) directly phosphorylate MLKL at Tyr376, promoting MLKL oligomerization but not its membrane translocation or RIPK3-mediated phosphorylation; TAM kinase inhibition or KO potently inhibits necroptotic death. Pharmacological inhibition, genetic KO, phospho-site mutagenesis, MLKL oligomerization assays, in vivo SIRS model Molecular cell High 31230815
2021 Conformational interconversion of the MLKL pseudokinase domain accompanies MLKL disengagement from RIPK3 following necroptotic stimulation; crystal structures of the pseudokinase domain in complex with Monobodies reveal two distinct conformations, and one Monobody epitope is only exposed after phosphorylated MLKL disengages from RIPK3. Crystal structure of MLKL pseudokinase domain–Monobody complexes, synthetic binding proteins, cell-based necroptosis assays Nature communications High 33850121
2023 RIPK3-mediated phosphorylation of the human MLKL activation loop drives pseudokinase domain dimerization, which nucleates elongated homotetramers via a central coiled coil formed by the ~80 Å brace helix; tetramerization is an essential prerequisite for release and reorganization of four-helix bundle domains for membrane permeabilization. Crystal structure, negative stain electron microscopy, mutagenesis, cell death assays Nature communications High 37884510
2021 MLKL kinase-like domain dimerization is an obligatory step following RIP3-induced phosphorylation; two inter-dimer interfaces identified in the crystal structure are required for RIP3-induced MLKL oligomerization and necroptosis. Subsequent self-assembly via an internal coiled-coil region forms the full oligomer. Crystal structure of human MLKL kinase-like domain, interface mutagenesis, oligomerization assays, cell death assays Cell death & disease High 34158471
2020 A missense mutation in the MLKL brace region (D139V in mouse) confers constitutive, RIPK3-independent killing activity, demonstrating that the brace helix connecting the 4HB and pseudokinase domains is a critical regulatory element that normally restrains MLKL activation. ENU mutagenesis screen, in vivo mouse model, RIPK3 KO epistasis, cell death assays Nature communications High 32561755
2016 MLKL 4HB domain function is evolutionarily divergent: mouse, horse, and frog 4HB domains induce cell death when expressed in murine fibroblasts, but human, chicken and stickleback 4HB domains cannot without forced dimerization. Nevertheless, recombinant 4HB proteins from all species permeabilize liposomes, preferentially those mimicking plasma membrane composition. Cross-species expression in murine fibroblasts, forced dimerization constructs, liposome permeabilization assays Cell death and differentiation High 26868910
2014 Necroptosis induced by RIPK3 requires MLKL but not Drp1; MLKL-deficient MEFs are completely protected from RIPK3-induced necroptosis (rescuable only by apoptosis inhibitor in WT), while Drp1-KO MEFs are not protected, and constitutively active MLKL mutants kill cells independently of Drp1. MLKL-KO MEFs, Drp1-KO MEFs, inducible RIPK3 dimerization system, constitutively active MLKL mutants, caspase inhibition Cell death & disease High 24577084
2017 Active MLKL triggers NLRP3 inflammasome activation in a cell-intrinsic manner, requiring: (i) the death effector 4HB domain, (ii) MLKL oligomerization and membrane association, and (iii) reduction in intracellular potassium concentration; this leads to ASC speck formation and caspase-1-dependent IL-1β cleavage before cell lysis. Domain mutagenesis, NLRP3/caspase-1/GSDMD KO cells, live imaging of ASC specks, K+ efflux measurement, chemogenetic MLKL activation Proceedings of the National Academy of Sciences of the United States of America High 28096356
2017 MLKL activation triggers potassium efflux and NLRP3 inflammasome assembly, which is required for IL-1β processing during necroptosis; MLKL-induced membrane disruption also allows IL-1β release independently of gasdermin-D. Chemogenetic MLKL activation system, NLRP3 KO, caspase-1 KO, GSDMD KO, K+ efflux measurement Journal of immunology High 28130493
2018 The pseudokinase MLKL activates PAD4-dependent NET formation in necroptotic neutrophils; MLKL translocates from cytoplasm to plasma membrane and stimulates downstream NADPH oxidase-independent ROS, nuclear membrane breakdown, chromatin decondensation, histone hypercitrullination, and NET extrusion. PAD4 acts downstream of RIPK1/RIPK3/MLKL. RIPK3 KO and MLKL KO neutrophils, PAD4 KO, live imaging, ROS measurement, NET quantification Science signaling High 30181240
2018 MLKL undergoes injury-induced phosphorylation at serine 441 in Schwann cells, distinct from the necroptosis-inducing RIP3-mediated phosphorylation, and this targets MLKL to the myelin sheath membrane to promote myelin breakdown after sciatic nerve injury; Schwann cell–specific Mlkl KO delays myelin breakdown and reduces nerve regeneration. Schwann cell-specific KO mouse, phospho-site specific analysis, in vivo nerve injury model, overexpression studies Molecular cell High 30344099
2021 RIPK3 and MLKL shuttle between nucleus and cytoplasm; during necroptosis, nuclear MLKL becomes phosphorylated and oligomerized, and pharmacological inhibition of nuclear export prevents cytosolic RIPK3/MLKL oligomerization and reduces necroptotic cell death. Nuclear/cytoplasmic fractionation, nuclear export inhibitors, immunofluorescence, cell death assays Communications biology Medium 30271893
2021 Oligomerization-induced multi-mono-ubiquitylation of MLKL occurs on at least four lysine residues in the membrane-associated fraction following MLKL activation; ubiquitylated MLKL is turned over via proteasome and lysosome; constitutive removal of ubiquitin by a MLKL-DUB fusion licenses MLKL auto-activation independent of necroptotic signaling. Mass spectrometry, MLKL-DUB fusion construct, membrane-targeted DUB, proteasome/lysosome inhibitors, KO cells The EMBO journal High 34698396
2022 K63-linked polyubiquitin chains conjugated to distinct lysine residues in the N-terminal HeLo domain of phosphorylated MLKL (facilitated by E3 ligase ITCH via WW domain interaction) divert MLKL to endosomes and extracellular vesicles instead of the plasma membrane, enhancing endosomal trafficking of intracellular bacteria to lysosomes. Site-directed ubiquitylation mutants, ITCH Co-IP/pulldown, bacterial infection assays, EV isolation Cell death and differentiation High 34999730
2020 Heat shock protein Hsp70 interacts with the N-terminal domain (NTD) of MLKL via its substrate-binding domain (SBD) and promotes MLKL NTD polymerization into amyloid-like structures; Hsp70 SBD is sufficient for this function, and NBC1 (which covalently conjugates Cys574/Cys603 of SBD) blocks MLKL polymerization and necroptosis. Hsp70 also stabilizes MLKL protein under basal conditions. In vitro pulldown, recombinant protein reconstitution of polymerization, site-directed mutagenesis (Cys574/Cys603), small-molecule inhibitor, Hsp70 knockdown Proceedings of the National Academy of Sciences of the United States of America High 32156734
2021 CAMK2/CaMKII phosphorylates MLKL in response to serum/amino acid starvation in a RIPK3-independent manner, and this promotes autophagic flux by facilitating autophagosome-lysosome fusion; this is mechanistically distinct from the necroptotic RIP3-MLKL axis. CAMK2 pharmacological inhibition and genetic KO, MLKL KO, autophagy reporter (mRFP-GFP-LC3), LC3-II and p62 assays Autophagy High 34282994
2023 MLKL forms amyloid-like polymers that translocate to lysosomal membranes during necroptosis, inducing lysosome clustering, fusion, and lysosomal membrane permeabilization (LMP), which releases cathepsins (especially Cathepsin B) into the cytosol, contributing to cell death. MLKL NTD expression constructs, lysosomal localization by imaging, LMP assay, cathepsin B knockdown/inhibition, cell death assays Cell death and differentiation High 37996483
2019 MLKL directly binds intracellular Listeria monocytogenes in the cytosol and inhibits bacterial replication in a cell death–independent manner; RIPK3-mediated phosphorylation of MLKL does not lead to host cell killing during Listeria infection. Co-localization microscopy, MLKL-bacteria binding assay, RIPK3 KO and MLKL KO cells, in vivo infection model The Journal of cell biology Medium 30975711
2019 The E3 ubiquitin ligase Skp2 interacts with MLKL and promotes its ubiquitination and proteasomal degradation; Skp2 overexpression in cisplatin-resistant NSCLC cells reduces MLKL levels, and Skp2 inhibition restores MLKL and sensitizes cells to necroptosis. Co-IP, ubiquitination assay, Skp2 KD/KO, in vivo xenograft model Communications biology Medium 37532777
2019 BRD4, IRF1, P-TEFb, and RNA polymerase II form a transcriptional complex at the MLKL promoter to regulate MLKL expression; BET inhibitors disrupt this complex and downregulate MLKL, suppressing necroptosis. ChIP, Co-IP of transcription complex, BET inhibitor treatment, IRF1 KO, MLKL rescue experiments Cell death and differentiation Medium 30644439
2020 MLKL-dependent (but RIP3-independent) signaling contributes to diet-induced liver injury by inhibiting autophagy; MLKL translocates first to autophagosomes and then to the plasma membrane in response to palmitic acid independently of Rip3, and Mlkl overexpression alone blocks autophagy. Mlkl-KO vs Rip3-KO mice on Western diet, mRFP-GFP-LC3 autophagy reporter in hepatocytes, subcellular fractionation, MLKL overexpression Journal of hepatology High 32220583
2025 Phosphorylated MLKL translocates not only to the plasma membrane but also to mitochondria, inducing microtubule-dependent release of mitochondrial DNA (mtDNA) into the cytosol, which activates the cGAS-STING pathway and upregulates IFN-β expression during necroptosis. Subcellular fractionation, live imaging, mtDNA quantification, cGAS-STING KO cells, in vivo IBD model with STING pathway interference Molecular cell High 40614706
2018 Interferon signaling (via cGAS/STING-driven constitutive IFN) maintains MLKL expression above a critical threshold required for MLKL oligomerization and necroptotic cell death in macrophages; it is the pre-established IFN status, not LPS-induced IFN, that is critical for early necroptosis initiation. cGAS/STING KO macrophages, MLKL expression rescue, oligomerization assays, IFN neutralization Cell death and differentiation Medium 29786074
2018 Interferons (type I and II) transcriptionally upregulate MLKL expression via IRF1 and STAT1; IFNγ increases MLKL mRNA and protein levels in a transcription-dependent manner, and IRF1 or STAT1 knockdown attenuates IFNγ-mediated MLKL induction. Actinomycin D chase, IRF1 KO, STAT1 KD, RT-PCR, Western blot Neoplasia Medium 30521981
2016 MLKL can be activated in experimental hepatitis through an MLKL-dependent pathway that is independent of RIPK3; IFN-γ induces MLKL expression via STAT1 transcription factor activation, connecting cytokine-driven inflammation to programmed necrosis. Genetic KO (MLKL-deficient vs RIPK3-deficient mice), pharmacological MLKL inhibition, STAT1 pathway analysis, human biopsy samples The Journal of clinical investigation High 27756058
2021 MLKL deficiency in myeloid cells (macrophages/Kupffer cells) impairs phagocytosis; LPS induces STAT1-mediated MLKL expression, phosphorylation, and translocation to intracellular compartments (phagosomes/lysosomes but not plasma membrane) in macrophages, and pharmacological or genetic MLKL inhibition suppresses phagocytic capacity. Bone marrow transplant chimeras (Mlkl-KO → WT and WT → Mlkl-KO), in vitro phagocytosis assay, subcellular fractionation, in vivo bioparticle uptake assay Hepatology High 35689613
2022 MLKL colocalizes with late endosomes/multivesicular bodies in macrophages handling atherogenic lipoproteins; Mlkl knockdown impairs lipid trafficking through multivesicular bodies, leading to enhanced lipid accumulation in macrophage foam cells. Antisense oligonucleotide knockdown in vivo, immunofluorescence co-localization with endosomal markers, in vitro foam cell assay Arteriosclerosis, thrombosis, and vascular biology Medium 32212851
2023 MLKL regulates ER-mitochondrial Mg2+ dynamics in RIPK3-deficient HCC cells; MLKL deficiency restricts ER Mg2+ release and mitochondrial Mg2+ uptake, causing ER dysfunction and mitochondrial oxidative stress, increasing susceptibility to metabolic stress-induced parthanatos (PARP1-dependent cell death). MLKL KO in HCC lines, Mg2+ live imaging, mitochondrial membrane potential assay, PARP inhibitor rescue, in vivo syngeneic tumor model Cell discovery Medium 36650126
2021 MLKL activates caspase-8-independent NLRP3 inflammasome activation and IL-1β secretion during influenza A virus infection in macrophages through potassium efflux; in the absence of MLKL, caspase-8 serves as a redundant mechanism for IL-1β maturation and inflammatory cell death. MLKL KO macrophages, NLRP3 inhibition, K+ efflux measurement, caspase-8 KO epistasis, in vivo IAV infection model mBio High 36852999

Source papers

Stage 0 corpus · 100 papers · ranked by NIH iCite citations
Year Title Journal Citations PMID
2014 Mixed lineage kinase domain-like protein MLKL causes necrotic membrane disruption upon phosphorylation by RIP3. Molecular cell 1319 24703947
2013 The pseudokinase MLKL mediates necroptosis via a molecular switch mechanism. Immunity 1036 24012422
2013 Toll-like receptor 3-mediated necrosis via TRIF, RIP3, and MLKL. The Journal of biological chemistry 802 24019532
2015 RIPK3 promotes cell death and NLRP3 inflammasome activation in the absence of MLKL. Nature communications 566 25693118
2017 ESCRT-III Acts Downstream of MLKL to Regulate Necroptotic Cell Death and Its Consequences. Cell 540 28388412
2014 RIPK1 regulates RIPK3-MLKL-driven systemic inflammation and emergency hematopoiesis. Cell 514 24813849
2017 Active MLKL triggers the NLRP3 inflammasome in a cell-intrinsic manner. Proceedings of the National Academy of Sciences of the United States of America 413 28096356
2017 MLKL, the Protein that Mediates Necroptosis, Also Regulates Endosomal Trafficking and Extracellular Vesicle Generation. Immunity 318 28666573
2020 MLKL trafficking and accumulation at the plasma membrane control the kinetics and threshold for necroptosis. Nature communications 277 32561730
2016 Cytotoxicity of crystals involves RIPK3-MLKL-mediated necroptosis. Nature communications 228 26817517
2016 Sequential Engagement of Distinct MLKL Phosphatidylinositol-Binding Sites Executes Necroptosis. Molecular cell 200 26853145
2017 MLKL Activation Triggers NLRP3-Mediated Processing and Release of IL-1β Independently of Gasdermin-D. Journal of immunology (Baltimore, Md. : 1950) 199 28130493
2015 PMA and crystal-induced neutrophil extracellular trap formation involves RIPK1-RIPK3-MLKL signaling. European journal of immunology 195 26531064
2018 Conformational switching of the pseudokinase domain promotes human MLKL tetramerization and cell death by necroptosis. Nature communications 162 29930286
2020 MLKL-dependent signaling regulates autophagic flux in a murine model of non-alcohol-associated fatty liver and steatohepatitis. Journal of hepatology 161 32220583
2021 ZBP1-MLKL necroptotic signaling potentiates radiation-induced antitumor immunity via intratumoral STING pathway activation. Science advances 152 34613770
2018 Constitutive interferon signaling maintains critical threshold of MLKL expression to license necroptosis. Cell death and differentiation 147 29786074
2016 Necrosome core machinery: MLKL. Cellular and molecular life sciences : CMLS 147 27048809
2016 Contribution of RIP3 and MLKL to immunogenic cell death signaling in cancer chemotherapy. Oncoimmunology 144 27471616
2018 Nuclear RIPK3 and MLKL contribute to cytosolic necrosome formation and necroptosis. Communications biology 135 30271893
2016 The pseudokinase MLKL mediates programmed hepatocellular necrosis independently of RIPK3 during hepatitis. The Journal of clinical investigation 134 27756058
2017 RIPK1/RIPK3/MLKL-mediated necroptosis contributes to compression-induced rat nucleus pulposus cells death. Apoptosis : an international journal on programmed cell death 116 28289909
2019 TAM Kinases Promote Necroptosis by Regulating Oligomerization of MLKL. Molecular cell 115 31230815
2022 GSK872 and necrostatin-1 protect retinal ganglion cells against necroptosis through inhibition of RIP1/RIP3/MLKL pathway in glutamate-induced retinal excitotoxic model of glaucoma. Journal of neuroinflammation 110 36289519
2019 Viral MLKL Homologs Subvert Necroptotic Cell Death by Sequestering Cellular RIPK3. Cell reports 103 31553902
2016 Evolutionary divergence of the necroptosis effector MLKL. Cell death and differentiation 103 26868910
2019 The pseudokinase MLKL regulates hepatic insulin sensitivity independently of inflammation. Molecular metabolism 91 30837196
2014 Necroptosis induced by RIPK3 requires MLKL but not Drp1. Cell death & disease 89 24577084
2018 The pseudokinase MLKL activates PAD4-dependent NET formation in necroptotic neutrophils. Science signaling 84 30181240
2021 Conformational interconversion of MLKL and disengagement from RIPK3 precede cell death by necroptosis. Nature communications 83 33850121
2020 A missense mutation in the MLKL brace region promotes lethal neonatal inflammation and hematopoietic dysfunction. Nature communications 83 32561755
2018 Mixed Lineage Kinase Domain-like Protein MLKL Breaks Down Myelin following Nerve Injury. Molecular cell 78 30344099
2020 The Killer Pseudokinase Mixed Lineage Kinase Domain-Like Protein (MLKL). Cold Spring Harbor perspectives in biology 74 31712266
2019 RIP1/RIP3/MLKL mediates dopaminergic neuron necroptosis in a mouse model of Parkinson disease. Laboratory investigation; a journal of technical methods and pathology 71 31506635
2017 Insane in the membrane: a structural perspective of MLKL function in necroptosis. Immunology and cell biology 70 27999433
2024 Gasdermin and MLKL necrotic cell death effectors: Signaling and diseases. Immunity 68 38479360
2021 CAMK2/CaMKII activates MLKL in short-term starvation to facilitate autophagic flux. Autophagy 67 34282994
2015 Upregulated RIP3 Expression Potentiates MLKL Phosphorylation-Mediated Programmed Necrosis in Toxic Epidermal Necrolysis. The Journal of investigative dermatology 66 25748555
2021 Oligomerization-driven MLKL ubiquitylation antagonizes necroptosis. The EMBO journal 64 34698396
2019 Necroptosis mediators RIPK3 and MLKL suppress intracellular Listeria replication independently of host cell killing. The Journal of cell biology 63 30975711
2022 Loss of MLKL ameliorates liver fibrosis by inhibiting hepatocyte necroptosis and hepatic stellate cell activation. Theranostics 59 35836819
2021 A toolbox for imaging RIPK1, RIPK3, and MLKL in mouse and human cells. Cell death and differentiation 59 33589776
2023 Dimethyl fumarate inhibits necroptosis and alleviates systemic inflammatory response syndrome by blocking the RIPK1-RIPK3-MLKL axis. Pharmacological research 58 36796462
2022 Necrosulfonamide ameliorates intestinal inflammation via inhibiting GSDMD-medicated pyroptosis and MLKL-mediated necroptosis. Biochemical pharmacology 58 36347275
2020 Loss of MLKL (Mixed Lineage Kinase Domain-Like Protein) Decreases Necrotic Core but Increases Macrophage Lipid Accumulation in Atherosclerosis. Arteriosclerosis, thrombosis, and vascular biology 58 32212851
2023 MLKL polymerization-induced lysosomal membrane permeabilization promotes necroptosis. Cell death and differentiation 56 37996483
2022 Caspase-8 auto-cleavage regulates programmed cell death and collaborates with RIPK3/MLKL to prevent lymphopenia. Cell death and differentiation 51 35064213
2021 The molecular mechanisms of MLKL-dependent and MLKL-independent necrosis. Journal of molecular cell biology 50 33064829
2019 Ptpn6 inhibits caspase-8- and Ripk3/Mlkl-dependent inflammation. Nature immunology 50 31819256
2019 MLKL contributes to shikonin-induced glioma cell necroptosis via promotion of chromatinolysis. Cancer letters 49 31560934
2018 Interferons Transcriptionally Up-Regulate MLKL Expression in Cancer Cells. Neoplasia (New York, N.Y.) 46 30521981
2023 A RIPK3-independent role of MLKL in suppressing parthanatos promotes immune evasion in hepatocellular carcinoma. Cell discovery 45 36650126
2023 MLKL-Driven Inflammasome Activation and Caspase-8 Mediate Inflammatory Cell Death in Influenza A Virus Infection. mBio 45 36852999
2015 Post-translational control of RIPK3 and MLKL mediated necroptotic cell death. F1000Research 44 27158445
2021 Surviving death: emerging concepts of RIPK3 and MLKL ubiquitination in the regulation of necroptosis. The FEBS journal 43 34710282
2018 RIPK1-RIPK3-MLKL-Associated Necroptosis Drives Leishmania infantum Killing in Neutrophils. Frontiers in immunology 43 30154785
2023 Macrophage-derived MLKL in alcohol-associated liver disease: Regulation of phagocytosis. Hepatology (Baltimore, Md.) 42 35689613
2023 The Many Faces of MLKL, the Executor of Necroptosis. International journal of molecular sciences 42 37373257
2020 Necroptosis-blocking compound NBC1 targets heat shock protein 70 to inhibit MLKL polymerization and necroptosis. Proceedings of the National Academy of Sciences of the United States of America 42 32156734
2023 Norovirus MLKL-like protein initiates cell death to induce viral egress. Nature 41 36991121
2019 Combined Knockout of RIPK3 and MLKL Reveals Unexpected Outcome in Tissue Injury and Inflammation. Frontiers in cell and developmental biology 37 30842945
2023 MLKL deficiency alleviates neuroinflammation and motor deficits in the α-synuclein transgenic mouse model of Parkinson's disease. Molecular neurodegeneration 36 38041169
2021 Differential role of MLKL in alcohol-associated and non-alcohol-associated fatty liver diseases in mice and humans. JCI insight 36 33616081
2021 RIPK3 Activates MLKL-mediated Necroptosis and Inflammasome Signaling during Streptococcus Infection. American journal of respiratory cell and molecular biology 35 33625952
2022 Site-specific ubiquitination of MLKL targets it to endosomes and targets Listeria and Yersinia to the lysosomes. Cell death and differentiation 34 34999730
2020 RIP1/RIP3/MLKL-mediated necroptosis contributes to vinblastine-induced myocardial damage. Molecular and cellular biochemistry 33 33247805
2019 MLKL attenuates colon inflammation and colitis-tumorigenesis via suppression of inflammatory responses. Cancer letters 33 31158430
2018 Necroptosis: MLKL Polymerization. Journal of nature and science 33 30294675
2024 Unveiling the interplay of MAPK/NF-κB/MLKL axis in brain health: Omega-3 as a promising candidates against copper neurotoxicity. Journal of environmental management 31 39357438
2021 The MLKL kinase-like domain dimerization is an indispensable step of mammalian MLKL activation in necroptosis signaling. Cell death & disease 31 34158471
2023 Necroptosis of macrophage is a key pathological feature in biliary atresia via GDCA/S1PR2/ZBP1/p-MLKL axis. Cell death & disease 30 36859525
2023 RIPK3-MLKL necroptotic signalling amplifies STING pathway and exacerbates lethal sepsis. Clinical and translational medicine 30 37475188
2022 Diet-derived ergothioneine induces necroptosis in colorectal cancer cells by activating the SIRT3/MLKL pathway. FEBS letters 30 35122251
2019 The bromodomain protein BRD4 positively regulates necroptosis via modulating MLKL expression. Cell death and differentiation 30 30644439
2023 Inhibition of gingival fibroblast necroptosis mediated by RIPK3/MLKL attenuates periodontitis. Journal of clinical periodontology 28 37366309
2023 Phosphorylation-dependent pseudokinase domain dimerization drives full-length MLKL oligomerization. Nature communications 28 37884510
2021 A family harboring an MLKL loss of function variant implicates impaired necroptosis in diabetes. Cell death & disease 28 33795639
2020 Cell-specific activation of RIPK1 and MLKL after intracerebral hemorrhage in mice. Journal of cerebral blood flow and metabolism : official journal of the International Society of Cerebral Blood Flow and Metabolism 28 33210566
2015 A tale of two domains - a structural perspective of the pseudokinase, MLKL. The FEBS journal 27 26337687
2023 Celastrol inhibits necroptosis by attenuating the RIPK1/RIPK3/MLKL pathway and confers protection against acute pancreatitis in mice. International immunopharmacology 26 37012867
2019 RIP1, RIP3, and MLKL Contribute to Cell Death Caused by Clostridium perfringens Enterotoxin. mBio 26 31848291
2025 MLKL activates the cGAS-STING pathway by releasing mitochondrial DNA upon necroptosis induction. Molecular cell 25 40614706
2023 Skp2-mediated MLKL degradation confers cisplatin-resistant in non-small cell lung cancer cells. Communications biology 25 37532777
2020 A novel neurodegenerative spectrum disorder in patients with MLKL deficiency. Cell death & disease 25 32358523
2023 Mitochondrial Trafficking of MLKL, Bak/Bax, and Drp1 Is Mediated by RIP1 and ROS which Leads to Decreased Mitochondrial Membrane Integrity during the Hyperglycemic Shift to Necroptosis. International journal of molecular sciences 24 37239951
2022 MLKL post-translational modifications: road signs to infection, inflammation and unknown destinations. Cell death and differentiation 24 36175538
2018 Non-Hematopoietic MLKL Protects Against Salmonella Mucosal Infection by Enhancing Inflammasome Activation. Frontiers in immunology 24 29456533
2023 Absence of Either Ripk3 or Mlkl Reduces Incidence of Hepatocellular Carcinoma Independent of Liver Fibrosis. Molecular cancer research : MCR 23 37204757
2023 PROTACs Targeting MLKL Protect Cells from Necroptosis. Journal of medicinal chemistry 23 37535857
2021 The Role of the Key Effector of Necroptotic Cell Death, MLKL, in Mouse Models of Disease. Biomolecules 23 34071602
2023 GSK2795039 prevents RIP1-RIP3-MLKL-mediated cardiomyocyte necroptosis in doxorubicin-induced heart failure through inhibition of NADPH oxidase-derived oxidative stress. Toxicology and applied pharmacology 22 36764612
2022 Interferon-γ Preferentially Promotes Necroptosis of Lung Epithelial Cells by Upregulating MLKL. Cells 22 35159372
2022 The role of MLKL in Hepatic Ischemia-Reperfusion Injury of Alcoholic Steatotic Livers. International journal of biological sciences 22 35173541
2022 Discovery of a New Class of Uracil Derivatives as Potential Mixed Lineage Kinase Domain-like Protein (MLKL) Inhibitors. Journal of medicinal chemistry 22 36136378
2020 RIPK3-MLKL-Mediated Neutrophil Death Requires Concurrent Activation of Fibroblast Activation Protein-α. Journal of immunology (Baltimore, Md. : 1950) 22 32796025
2021 MLKL and CaMKII Are Involved in RIPK3-Mediated Smooth Muscle Cell Necroptosis. Cells 21 34572045
2022 RIPK3-MLKL signaling activates mitochondrial CaMKII and drives intrarenal extracellular matrix production during CKD. Matrix biology : journal of the International Society for Matrix Biology 20 35964866
2024 MLKL-mediated endothelial necroptosis drives vascular damage and mortality in systemic inflammatory response syndrome. Cellular & molecular immunology 19 39349742
2022 Macrophage Infiltration Initiates RIP3/MLKL-Dependent Necroptosis in Paclitaxel-Induced Neuropathic Pain. Mediators of inflammation 19 36164389
2022 1,3-Dichloro-2-propanol-Induced Renal Tubular Cell Necroptosis through the ROS/RIPK3/MLKL Pathway. Journal of agricultural and food chemistry 18 36000575