A5051709970- Cell death mechanisms and regulation
- interferon and immune responses
- Advanced Proteomics Techniques and Applications
- CRISPR and Genetic Engineering
- Phagocytosis and Immune Regulation
- Mass Spectrometry Techniques and Applications
- Metabolomics and Mass Spectrometry Studies
- Cancer-related Molecular Pathways
- Psoriasis: Treatment and Pathogenesis
- Cytokine Signaling Pathways and Interactions
- SARS-CoV-2 and COVID-19 Research
- PARP inhibition in cancer therapy
- Protein Kinase Regulation and GTPase Signaling
- Immune Response and Inflammation
- DNA Repair Mechanisms
- Protein Structure and Dynamics
- RNA Interference and Gene Delivery
- COVID-19 Clinical Research Studies
- Microtubule and mitosis dynamics
- RNA regulation and disease
- Urticaria and Related Conditions
- Glycosylation and Glycoproteins Research
- RNA and protein synthesis mechanisms
- NF-κB Signaling Pathways
- Ubiquitin and proteasome pathways
Max Planck Institute of Biochemistry
2019-2025
Walter and Eliza Hall Institute of Medical Research
2014-2025
The University of Melbourne
2013-2025
Institute for Molecular Science
2023
La Trobe University
2023
Centre for Inflammation Research
2023
King's College London
2023
Duke University
2023
Max Planck Society
2019
Innsbruck Medical University
2012-2015
Significance The four-helix bundle (4HB) domain of Mixed Lineage Kinase Domain-Like (MLKL) bears two clusters residues that are required for cell death by necroptosis. Mutations within a cluster centered on the α4 helix 4HB MLKL prevented its membrane translocation, oligomerization, and ability to induce This is composed principally acidic therefore challenges idea engages negatively charged phospholipid membranes via conventional positively interaction surface. importance translocation...
Humans encode two inflammatory caspases that detect cytoplasmic LPS, caspase‐4 and caspase‐5. When activated, these trigger pyroptotic cell death caspase‐1‐dependent IL‐1β production; however the mechanism underlying this process is not yet confirmed. We now show a specific NLRP3 inhibitor, MCC950, prevents caspase‐4/5‐dependent production elicited by transfected LPS. Given both caspase‐5 can it possible proteins exhibit some degree of redundancy. Therefore, we generated human monocytic...
Necroptotic cell death is mediated by the most terminal known effector of pathway, MLKL. Precisely how phosphorylation MLKL pseudokinase domain activation loop upstream kinase, RIPK3, induces unmasking N-terminal executioner four-helix bundle (4HB) MLKL, higher-order assemblies, and permeabilization plasma membranes remains poorly understood. Here, we reveal existence a basal monomeric conformer present in human cells prior to exposure necroptotic stimulus. Following activation, toggling...
The recent revolution in computational protein structure prediction provides folding models for entire proteomes, which can now be integrated with large-scale experimental data. Mass spectrometry (MS)-based proteomics has identified and quantified tens of thousands posttranslational modifications (PTMs), most them uncertain functional relevance. In this study, we determine the structural context these PTMs investigate how information leveraged to pinpoint potential regulatory sites. Our...
Abstract Toxic epidermal necrolysis (TEN) is a fatal drug-induced skin reaction triggered by common medications and an emerging public health issue 1–3 . Patients with TEN undergo severe sudden detachment caused keratinocyte cell death. Although molecular mechanisms that drive death have been proposed, the main drivers remain unknown, there no effective therapy for 4–6 Here, to systematically map changes are associated identify potential druggable targets, we utilized deep visual proteomics,...
Resistance to chemotherapy is a major problem in cancer treatment, and it frequently associated with failure of tumor cells undergo apoptosis. Birinapant, clinical SMAC mimetic, had been designed mimic the interaction between inhibitor apoptosis proteins (IAPs) SMAC/Diablo, thereby relieving IAP-mediated caspase inhibition promoting cells. We show that acute myeloid leukemia (AML) are sensitive birinapant-induced death emricasan/IDN-6556 augments, rather than prevents, killing by birinapant....
Abstract Necroptosis is a caspase-independent form of regulated cell death that has been implicated in the development range inflammatory, autoimmune and neurodegenerative diseases. The pseudokinase, Mixed Lineage Kinase Domain-Like (MLKL), most terminal known obligatory effector necroptosis pathway, activated following phosphorylation by Receptor Interacting Protein Kinase-3 (RIPK3). Activated MLKL translocates to membranes, leading membrane destabilisation subsequent death. However,...
Summary The global emergence of SARS-CoV-2 urgently requires an in-depth understanding molecular functions viral proteins and their interactions with the host proteome. Several individual omics studies have extended our knowledge COVID-19 pathophysiology 1–10 . Integration such datasets to obtain a holistic view virus-host define pathogenic properties is limited by heterogeneity experimental systems. We therefore conducted concurrent multi-omics study SARS-CoV. Using state-of-the-art...
The pseudokinase MLKL (mixed lineage kinase domain-like), has recently emerged as a critical component of the necroptosis cell death pathway. Although it is clear that phosphorylation activation loop in domain by upstream protein RIPK3 (receptor-interacting kinase-3), crucial to trigger activation, remained unclear whether other events modulate function. By reconstituting Mlkl−/−, Ripk3−/− and Mlkl−/−Ripk3−/− cells with phospho-site mutants, we compared function known sites regulating three...
The pseudokinase MLKL (mixed lineage kinase domain-like) was identified recently as an essential checkpoint in the programmed necrosis or ‘necroptosis’ cell death pathway. In present study, we report crystal structure of human domain at 1.7 Å (1 Å=0.1 nm) resolution and probe its nucleotide-binding mechanism by performing structure-based mutagenesis. By comparing structures determinants mouse orthologues, study provides insights into evolution mechanisms among pseudokinases their mechanistic...
Abstract Protein ubiquitination is involved in virtually all cellular processes. Enrichment strategies employing antibodies targeting ubiquitin-derived diGly remnants combined with mass spectrometry (MS) have enabled investigations of ubiquitin signaling at a large scale. However, so far the power data independent acquisition (DIA) regards to sensitivity single run analysis and completeness not yet been explored. Here, we develop sensitive workflow combining antibody-based enrichment...
Abstract MLKL is the essential effector of necroptosis, a form programmed lytic cell death. We have isolated mouse strain with single missense mutation, Mlkl D139V , that alters two-helix ‘brace’ connects killer four-helix bundle and regulatory pseudokinase domains. This confers constitutive, RIPK3 independent killing activity to MLKL. Homozygous mutant mice develop lethal postnatal inflammation salivary glands mediastinum. The normal embryonic development homozygotes until birth, absence...
Bacterial cell wall components provide various unique molecular structures that are detected by pattern recognition receptors (PRRs) of the innate immune system as non-self. Most bacterial species form a consists peptidoglycan (PGN), polymeric structure comprising alternating amino sugars strands cross-linked short peptides. Muramyl dipeptide (MDP) has been well documented minimal immunogenic component peptidoglycan1-3. MDP is sensed cytosolic nucleotide-binding oligomerization...
TRAF2 is a component of TNF superfamily signalling complexes and plays an essential role in the regulation homeostasis immune cells. deficient mice die around birth, therefore its adult tissues not well-explored. Furthermore, RING controversial. It has been claimed that atypical cannot function as ubiquitin E3 ligase but counterclaimed requires co-factor, sphingosine-1-phosphate, generated by enzyme sphingosine kinase 1, to ligase. Keratinocyte-specific deletion Traf2, Sphk1 deficiency,...
The inflammatory functions of the cytokine tumor necrosis factor (TNF) rely on its ability to induce production and cell death. Caspase-dependent caspase-independent pathways-apoptosis necroptosis, respectively-regulate immunogenicity by release distinct sets cellular proteins. To obtain an unbiased, systems-level understanding this important process, we here applied mass spectrometry-based proteomics dissect protein during apoptosis necroptosis. We report hundreds proteins released from...
Abstract Tumor necrosis factor (TNF) is one of the few cytokines successfully targeted by therapies against inflammatory diseases. However, blocking this well studied and pleiotropic ligand can cause dramatic side-effects. Here, we reason that a systems-level proteomic analysis TNF signaling could dissect its diverse functions offer base for developing more therapies. Therefore, combine phosphoproteomics time course experiments with subcellular localization kinase inhibitor to identify...
Fibrosis represents the uncontrolled replacement of parenchymal tissue with extracellular matrix (ECM) produced by myofibroblasts. While genetic fate-tracing and single-cell RNA-Seq technologies have helped elucidate fibroblast heterogeneity ontogeny beyond to myofibroblast differentiation, newly identified populations remain ill defined, respect both molecular cues driving their differentiation subsequent role in fibrosis. Using an unbiased approach, we metalloprotease ADAMTS12 as a...