A5009119703- Immune Cell Function and Interaction
- Streptococcal Infections and Treatments
- Lipid Membrane Structure and Behavior
- Autoimmune and Inflammatory Disorders Research
- Bacteriophages and microbial interactions
- Force Microscopy Techniques and Applications
- Inflammasome and immune disorders
- Nanopore and Nanochannel Transport Studies
- Antimicrobial Resistance in Staphylococcus
- CAR-T cell therapy research
- Phagocytosis and Immune Regulation
- Immunotherapy and Immune Responses
- ATP Synthase and ATPases Research
- RNA Interference and Gene Delivery
- Infective Endocarditis Diagnosis and Management
- Cellular Mechanics and Interactions
- Advanced Biosensing Techniques and Applications
- Amoebic Infections and Treatments
- Signaling Pathways in Disease
- T-cell and B-cell Immunology
- Lysosomal Storage Disorders Research
- Calcium signaling and nucleotide metabolism
- Cell Image Analysis Techniques
- Monoclonal and Polyclonal Antibodies Research
- Peptidase Inhibition and Analysis
Peter MacCallum Cancer Centre
2020-2025
The University of Melbourne
2024
London Centre for Nanotechnology
2014-2020
University College London
2014-2020
Institute of Structural and Molecular Biology
2016-2020
Imperial College London
2016
ETH Zurich
2013
Membrane attack complex/perforin/cholesterol-dependent cytolysin (MACPF/CDC) proteins constitute a major superfamily of pore-forming that act as bacterial virulence factors and effectors in immune defence. Upon binding to the membrane, they convert from soluble monomeric form oligomeric, membrane-inserted pores. Using real-time atomic force microscopy (AFM), electron (EM), structure fitting, we have mapped assembly pathways CDC unprecedented detail accuracy, focussing on suilysin...
Abstract The membrane attack complex (MAC) is a hetero-oligomeric protein assembly that kills pathogens by perforating their cell envelopes. MAC formed sequential of soluble complement proteins C5b, C6, C7, C8 and C9, but little known about the rate-limiting steps in this process. Here, we use rapid atomic force microscopy (AFM) imaging to show oligomerize within membrane, unlike structurally homologous bacterial pore-forming toxins. C5b-7 interacts with lipid bilayer prior recruiting C8. We...
Abstract Macrophage-expressed gene 1 (MPEG1/Perforin-2) is a perforin-like protein that functions within the phagolysosome to damage engulfed microbes. MPEG1 thought form pores in target membranes, however, its mode of action remains unknown. We use cryo-Electron Microscopy (cryo-EM) determine 2.4 Å structure hexadecameric assembly displays expected features soluble prepore complex. further discover prepore-like assemblies can be induced perforate membranes through acidification, such as...
Abstract Killer T cells (cytotoxic lymphocytes, CTLs) maintain immune homoeostasis by eliminating virus-infected and cancerous cells. CTLs achieve this forming an immunological synapse with their targets secreting a pore-forming protein (perforin) pro-apoptotic serine proteases (granzymes) into the synaptic cleft. Although CTL target cell are both exposed to perforin within synapse, only membrane is disrupted, while invariably spared. How escape unscathed remains mystery. Here, we report...
Abstract Cytotoxic lymphocytes are crucial to our immune system, primarily eliminating virus-infected or cancerous cells via perforin/granzyme killing. Perforin forms transmembrane pores in the plasma membrane, allowing granzymes enter target cell cytosol and trigger apoptosis. The prowess of cytotoxic efficiently eradicate has been widely harnessed immunotherapies against haematological cancers. Despite efforts achieve a similar outcome solid tumours, immunosuppressive acidic tumour...
Perforin is a pore forming protein used by cytotoxic T lymphocytes to remove cancerous or virus-infected cells during the immune response. During response, lymphocyte membrane becomes refractory perforin function accumulating densely ordered lipid rafts and externalizing negatively charged species. The dense packing lowers capacity of bind, lipids scavenge any residual before formation. Using atomic force microscopy on model systems, we here provide insight into molecular basis specificity.
Macrophage Expressed Gene-1 (MPEG-1; also termed Perforin-2) is an endosomal / phagolysosomal perforin-like protein that conserved across the metazoan kingdom and functions within phagolysosome to damage engulfed microbes. Like Membrane Attack Complex perforin, MPEG-1 has been postulated form pores in target membranes, however, its mode of action remains be established. We used single particle cryo-Electron Microscopy determine 2.4 Å structure a hexadecameric assembly displays expected...
Recently developed small-molecule inhibitors of the lysosomal protease dipeptidyl peptidase 1 (DPP1), also known as cathepsin C (CatC), can suppress suppurative inflammation in vivo by blocking processing zymogenic (pro-) forms neutrophil serine proteases (NSPs), including elastase, proteinase 3, and G. DPP1 plays an important role activating granzyme that are expressed cytotoxic T lymphocytes (CTL) natural killer (NK) cells. Therefore, it is critical to determine whether inhibition cause...
Abstract Cytotoxic lymphocytes are pivotal effectors in our immune system. Their most potent way to delete virus infected or cancerous target cells is by perforin/granzyme dependent killing. Perforin secreted into the immunological synapse and forms transmembrane pores cell plasma membrane, allowing granzymes enter cytosol trigger apoptosis. The prowess of cytotoxic efficiently eradicate has been widely harnessed immunotherapies against haematological cancers. Despite efforts achieve a...
Abstract The membrane attack complex (MAC) is a hetero-oligomeric protein assembly that kills pathogens by perforating their cell envelopes. MAC formed sequential of soluble complement proteins C5b, C6, C7, C8 and C9, but little known about the rate-limiting steps in this process. Here, we use rapid atomic force microscopy (AFM) imaging to show oligomerize within membrane, unlike structurally homologous bacterial pore-forming toxins. C5b6 interacts with lipid bilayer prior recruiting C7 C8....
Abstract A crucial point in making treatment decisions for cancer patients is the assessment of tumor aggressiveness. Currently, breast cancer, established prognostic markers exist that are routinely assessed by standard pathological examination. However, these parameters often not sufficient to stratify patients, especially those with early stages and adjuvant therapy frequently administered who might have been cured surgery anti-hormonal alone. The goal avoid over- under-treatment has led...
Abstract Perforin is a pore forming protein used by cytotoxic T lymphocytes to remove cancerous or virus-infected cells during immune response. During the response, lymphocyte membrane becomes refractory perforin function accumulating densely ordered lipid rafts and externalizing negatively charged species. The dense packing lowers capacity of bind, lipids scavenge any residual before formation. Using atomic force microscopy on model systems, we here provide insight into molecular basis specificity.