A5071161068- HIV Research and Treatment
- Parkinson's Disease Mechanisms and Treatments
- HIV/AIDS drug development and treatment
- Alzheimer's disease research and treatments
- Bacteriophages and microbial interactions
- Biotin and Related Studies
- Neurological disorders and treatments
- Botulinum Toxin and Related Neurological Disorders
- HIV/AIDS Research and Interventions
- interferon and immune responses
- Signaling Pathways in Disease
- Monoclonal and Polyclonal Antibodies Research
- Cytomegalovirus and herpesvirus research
- Advanced Biosensing Techniques and Applications
- Advanced biosensing and bioanalysis techniques
- RNA Research and Splicing
- Prion Diseases and Protein Misfolding
UNSW Sydney
2016-2024
EMBL Australia
2016-2024
ARC Centre of Excellence in Advanced Molecular Imaging
2018-2022
Uncoating of the metastable HIV-1 capsid is a tightly regulated disassembly process required for release viral cDNA prior to nuclear import. To understand intrinsic pathway and how it can be modulated, we have developed single-particle fluorescence microscopy method follow real-time uncoating kinetics authentic HIV capsids in vitro immediately after permeabilizing membrane. Opening first defect lattice rate-limiting step uncoating, which followed by rapid, catastrophic collapse. The...
The aggregation of alpha-synuclein (α-SYN) follows a cascade oligomeric, prefibrillar and fibrillar forms, culminating in the formation Lewy Bodies (LB), pathological hallmarks Parkinson's Disease. Although LB contain over 70 proteins, potential for interactions along pathway α-SYN is unknown. Here we propose map 65 proteins against different species α-SYN. We measured binding to monomeric using AlphaScreen, sensitive nano-bead luminescence assay detection protein interactions. To access...
α-Synuclein aggregation is a hallmark of Parkinson's disease and promising biomarker for early detection assessment progression. The prospect molecular test materializing with the recent developments methods based on amplification synuclein seeds (e.g. RT-QuIC or PMCA). Here we adapted single-molecule counting α-synuclein aggregates in cerebrospinal fluid (CSF), using simple 3D printed microscope. Single-molecule enable to probe events process used precise ThT-positive aggregates....
Abstract α‐Synuclein aggregation is a hallmark of Parkinson's disease and promising biomarker for early detection assessment progression. The prospect molecular test materializing with the recent developments methods based on amplification synuclein seeds (e.g. RT‐QuIC or PMCA). Here we adapted single‐molecule counting α‐synuclein aggregates in cerebrospinal fluid (CSF), using simple 3D printed microscope. Single‐molecule enable to probe events process used precise ThT‐positive aggregates....
The quantification of α-synuclein aggregates has emerged as a promising biomarker for synucleinopathies. Assays that amplify and detect such have revealed the presence seeding-competent species in biosamples patients diagnosed with Parkinson's disease. However, multiple species, oligomers amyloid fibrils, are formed during aggregation α-synuclein; these likely to coexist biological samples, thus it remains unclear which species(s) contributing signal detected seeding assays. To identify...
Efficient HIV-1 replication depends on interaction of the viral capsid with host protein cyclophilin A (CypA). CypA, a peptidylprolyl isomerase, binds to an exposed loop in CA via enzyme's active site. Recent structural analysis CypA complex tubes conjunction molecular dynamics simulations identified secondary binding site that allows bridging two hexameric subunits assembled lattice, leading stabilization (Liu et al. Nat Commun 7:10714, 2016). We performed mutational residues have been...
The human immunodeficiency virus 1 (HIV-1) capsid serves as a binding platform for proteins and small molecules from the host cell that regulate various steps in life cycle. However, there are currently no quantitative methods use assembled lattices to measure host–pathogen interaction dynamics. Here we developed single-molecule fluorescence biosensor using self-assembled tubes biorecognition elements imaged binders total internal reflection microscopy microfluidic setup. method is highly...
The stability of the HIV-1 capsid and spatiotemporal control its disassembly, a process called uncoating, need to be finely tuned for infection proceed. Biochemical methods measuring lattice disassembly in bulk are unable resolve intermediates uncoating reaction. We have developed single-particle fluorescence microscopy method follow real-time kinetics authentic HIV capsids vitro. assay utilizes immobilized viral particles that permeabilized with pore-former protein, is designed (1) detect...
The human immunodeficiency virus (HIV) capsid is a cone-shaped capsule formed from the viral protein (CA), which arranged into lattice of hexamers and pentamers. comprises multiple binding interfaces for recruitment host proteins macromolecules used by to establish infection. Here, we coassembled CA engineered pentamer cross-linking fluorescence labeling, spherical particles. spheres, resemble pentamer-rich structure end caps native HIV capsid, were immobilized onto surfaces as...
The HIV capsid is a multifunctional protein capsule that mediates the delivery of viral genetic material into nucleus target cell. Host cell proteins bind to number repeating binding sites on regulate steps in replication cycle. Here, we develop fluorescence fluctuation spectroscopy method using self-assembled particles as bait screen for fluorescence-labeled capsid-binding analytes ("prey" molecules) solution. assay capitalizes property multivalent interaction platform, facilitating high...
ABSTRACT Alpha-synuclein (αSyn) aggregates, detected in the biofluids of patients with Parkinson’s disease, have ability to catalyze their own aggregation, leading an increase number and size aggregates. This self-templated amplification is used by newly developed assays diagnose disease turned presence αSyn aggregates into a biomarker disease. It has become evident that can form fibrils slightly different structures, called “strains” or polymorphs, but little known about differential...
α-Synuclein (αSyn) aggregates, detected in the biofluids of patients with Parkinson's disease (PD), have ability to catalyze their own aggregation, leading an increase number and size aggregates. This self-templated amplification is used by newly developed assays diagnose turns presence αSyn aggregates into a biomarker disease. It has become evident that can form fibrils slightly different structures, called "strains" or polymorphs, but little known about differential reactivity diagnostic...
The innate immune system uses a multitude of receptors such as Toll‐like Receptors and Nod‐like to sense pathogens danger signals. Upon their activation, signalling cascade is initiated culminating in the transcription translation cytokines other proteins vital for host protection. Recent studies have discovered that an array these intracellular assemble into oligomeric platforms, consequently amplifying transmission signal from stimulated receptor activation relevant machinery....
The HIV capsid is a multifunctional protein capsule for delivery of the viral genetic material into nucleus target cell. Host cell proteins bind to number repeating binding sites on regulate steps in replication cycle. Here we develop fluorescence fluctuation spectroscopy method using self-assembled particles as bait screen fluorescence-labelled capsid-binding analytes (‘prey’ molecules) solution. assay capitalizes property multivalent interaction platform, facilitating high sensitivity...
The human immunodeficiency virus 1 (HIV-1) consists of a viral membrane surrounding the conical capsid. capsid is protein container assembled from approximately 1,500 copies (CA), functioning as reaction and transport chamber for genome after cell entry. Transmission electron microscopy (TEM) widely used technique characterizing ultrastructure isolated capsids removal membrane, which otherwise hinders negative staining structures inside particle TEM. Here, we provide protocol to permeabilize...
Abstract The quantification of α-synuclein (α-syn) aggregates has emerged as a promising biomarker for synucleinopathies. Assays that amplify and detect such have revealed the presence seeding-competent species in biosamples patients diagnosed with Parkinson’s disease. However, multiple oligomers amyloid fibrils, are formed during aggregation these likely to co-exist biological samples thus it remains unclear which species(s) contributing signal detected seeding assays. To identify can be...
Abstract The human immunodeficiency virus 1 (HIV-1) capsid serves as a binding platform for proteins and small molecules from the host cell that regulate various steps in life cycle. However, there are currently no quantitative methods use assembled lattices measuring host-pathogen interaction dynamics. Here we developed single molecule fluorescence biosensor using self-assembled tubes biorecognition elements imaged binders total internal reflection microscopy microfluidic setup. method is...
SUMMARY Cyclic GMP-AMP synthase (cGAS) is a primary sensor of aberrant DNA that governs an innate immune signaling cascade, leading to the induction type-I interferon response. We have previously identified polyglutamine binding protein 1, PQBP1, as adaptor molecule required for cGAS-mediated response lentiviruses, including human immunodeficiency virus 1 (HIV-1), but dispensable recognition viruses. HIV-1- encoded synthesized single copy from its RNA genome, and subsequently integrated into...