A5042822683- Alzheimer's disease research and treatments
- Advanced Fluorescence Microscopy Techniques
- Force Microscopy Techniques and Applications
- Parkinson's Disease Mechanisms and Treatments
- Electrochemical Analysis and Applications
- Advanced biosensing and bioanalysis techniques
- Analytical Chemistry and Sensors
- Monoclonal and Polyclonal Antibodies Research
- Microfluidic and Bio-sensing Technologies
- Prion Diseases and Protein Misfolding
- DNA and Nucleic Acid Chemistry
- Advanced Biosensing Techniques and Applications
- Lipid Membrane Structure and Behavior
- Spectroscopy and Quantum Chemical Studies
- Computational Drug Discovery Methods
- Microfluidic and Capillary Electrophoresis Applications
- Neuroinflammation and Neurodegeneration Mechanisms
- Advanced Electron Microscopy Techniques and Applications
- Protein Structure and Dynamics
- Molecular Junctions and Nanostructures
- Acoustic Wave Resonator Technologies
- RNA Interference and Gene Delivery
- Nanofabrication and Lithography Techniques
- T-cell and B-cell Immunology
- Immune Response and Inflammation
University of Cambridge
2016-2025
UK Dementia Research Institute
2017-2025
Addenbrooke's Hospital
2024
Sea Level Research (United Kingdom)
2024
Boehringer Ingelheim (United States)
2022
University College London
2006-2018
National Hospital for Neurology and Neurosurgery
2018
Bridge University
2012
University of Würzburg
2012
Universidad de Granada
2010
Here, we use single-molecule techniques to study the aggregation of α-synuclein, protein whose misfolding and deposition is associated with Parkinson's disease. We identify a conformational change from initially formed oligomers stable, more compact proteinase-K-resistant as key step that leads ultimately fibril formation. The result structural conversion generate much higher levels oxidative stress in rat primary neurons than do initially, showing they are damaging cells. remarkably slow,...
Reversible phase separation underpins the role of FUS in ribonucleoprotein granules and other membrane-free organelles is, part, driven by intrinsically disordered low-complexity (LC) domain FUS. Here, we report that cooperative cation-π interactions between tyrosines LC arginines structured C-terminal domains also contribute to separation. These are modulated post-translational arginine methylation, wherein hypomethylation strongly promotes gelation. Indeed, significant hypomethylation,...
We describe the isolation and detailed structural characterization of stable toxic oligomers α-synuclein that have accumulated during process amyloid formation. Our approach has allowed us to identify distinct subgroups probe their molecular architectures by using cryo-electron microscopy (cryoEM) image reconstruction techniques. Although exist in a range sizes, with different extents nature β-sheet content exposed hydrophobicity, they all possess hollow cylindrical architecture similarities...
Protein aggregation causes α-synuclein to switch from its physiological role a pathological toxic gain of function. Under conditions, monomeric improves ATP synthase efficiency. Here, we report that monomers generates beta sheet-rich oligomers localise the mitochondria in close proximity several mitochondrial proteins including synthase. Oligomeric impairs complex I-dependent respiration. Oligomers induce selective oxidation subunit and lipid peroxidation. These events increase probability...
Abstract Protein aggregation is a complex process resulting in the formation of heterogeneous mixtures aggregate populations that are closely linked to neurodegenerative conditions, such as Alzheimer’s disease. Here, we find soluble aggregates formed at different stages amyloid beta (Aβ42) induce disruption lipid bilayers and an inflammatory response extents. Further, by using gradient ultracentrifugation assay, show smaller those most potent inducing membrane permeability effectively...
Protein aggregation and oxidative stress are both key pathogenic processes in Parkinson's disease, although the mechanism by which misfolded proteins induce neuronal death remains unknown. In this study, we describe how of alpha-synuclein (α-S) from its monomeric form to soluble oligomeric state results aberrant free radical production toxicity.We first demonstrate excessive a human induced pluripotent stem-derived α-S triplication model at basal levels on application picomolar doses...
Double take: Double-barrel carbon nanoprobes with integrated distance control for simultaneous nanoscale electrochemical and ion conductance microscopy can be fabricated a wide range of probe sizes in less than two minutes. The allow noncontact topographical (left image) imaging (right) living neurons, as well localized K+ delivery neurotransmitter detection. Detailed facts importance to specialist readers are published "Supporting Information". Such documents peer-reviewed, but not...
Far-field optical microscopy using focused light is an important tool in a number of scientific disciplines including chemical, (bio)physical and biomedical research, particularly with respect to the study living cells organisms. Unfortunately, applicability microscope limited, since diffraction imposes limitations on spatial resolution image. Consequently details of, for example, cellular protein distributions, can be visualized only certain extent. Fortunately, recent years have witnessed...
The measurement of key molecules in individual cells with minimal disruption to the biological milieu is next frontier single-cell analyses. Nanoscale devices are ideal analytical tools because their small size and potential for high spatial temporal resolution recordings. Here, we report fabrication disk-shaped carbon nanoelectrodes whose radius can be precisely tuned within range 5-200 nm. functionalization nanoelectrode platinum allowed monitoring oxygen consumption outside inside a brain...
We report a microfluidic droplet-based approach enabling the measurement of chemical reactions individual enzyme molecules and its application to single-molecule-counting immunoassay. A device is used generate manipulate <10 fL droplets at rates up 1.3 × 10(6) per second, about 2 orders magnitude faster than has previously been reported. The femtodroplets produced with this can be encapsulate single biomolecular complexes tagged reporter enzyme; their small volume enables fluorescent product...
Significance Growing experimental evidence suggests that the pathological spreading of alpha-synuclein aggregates in Parkinson’s disease is mediated through a process templated seeding whereby catalyze conversion soluble protein molecules into their aggregated forms. A molecular-level understanding this still lacking. Here, we determine concentrations and numbers necessary for effective alpha-synuclein, thus providing quantitative framework to understand conditions when its seeded...
Abstract Protein aggregation and abnormal lipid homeostasis are both implicated in neurodegeneration through unknown mechanisms. Here we demonstrate that aggregate-membrane interaction is critical to induce a form of cell death called ferroptosis. Importantly, the drives ferroptosis depends on conformational structure aggregate, as well oxidation state membrane. We generated human stem cell-derived models synucleinopathy, characterized by intracellular formation α-synuclein aggregates bind...
Abstract Protein aggregation plays a key role in neurodegenerative disease, giving rise to small oligomers that may become cytotoxic cells. The fundamental microscopic reactions taking place during aggregation, and their rate constants, have been difficult determine due lack of suitable methods identify follow the low concentration over time. Here we use single-molecule fluorescence study repeat domain tau (K18), two mutant forms linked with familial frontotemporal dementia, deletion ΔK280...
Abstract Super-resolution microscopy allows biological systems to be studied at the nanoscale, but has been restricted providing only positional information. Here, we show that it is possible perform multi-dimensional super-resolution imaging determine both position and environmental properties of single-molecule fluorescent emitters. The method presented here exploits solvatochromic fluorogenic nile red extract emission spectrum each dye molecule simultaneously enabling mapping...
Aggregation of α-synuclein leads to the formation oligomeric intermediates that can interact with membranes form pores. However, it is unknown how this cell toxicity in Parkinson's disease. We investigated species-specific effects on Ca(2+) signalling primary neurons and astrocytes using live neuronal imaging electrophysiology artificial membranes. demonstrate induces an increase basal intracellular its unfolded monomeric state as well state. Electrophysiology demonstrated monomers induce...
Significance Protein oligomers are a fundamental component of amyloidogenic disorders such as Alzheimer’s disease, being both critical intermediates in the formation amyloid fibrils and most toxic species on protein aggregation pathway. Here, we derive general analytical expressions for chemical kinetics generated during amyloid-formation reactions explain how these may be used to categorize according their basic physicochemical properties. By applying this framework experimental kinetic...
Chaperone regulation of individual microscopic events in Aβ42 aggregation reveals the nature active sites on amyloid fibrils.
Abstract Aggregation of alpha-synuclein (α-Syn) drives Parkinson’s disease (PD), although the initial stages self-assembly and structural conversion have not been directly observed inside neurons. In this study, we tracked intracellular conformational states α-Syn using a single-molecule Förster resonance energy transfer (smFRET) biosensor, show here that converts from monomeric state into two distinct oligomeric in neurons concentration-dependent sequence-specific manner. Three-dimensional...
We have investigated the structure and unfolding kinetics of human telomeric intramolecular G quadruplex by using single-molecule fluorescence resonance energy transfer. An exploration conformational heterogeneity revealed two stable folded conformations, in both sodium- potassium-containing buffers, with small differences between their enthalpies entropies. Both conformations can be opened addition a 21-base complementary DNA oligonucleotide. The substates occurs at same rate, which showed...