A5091821729- Protein Structure and Dynamics
- Alzheimer's disease research and treatments
- Enzyme Structure and Function
- Heat shock proteins research
- Prion Diseases and Protein Misfolding
- Bacterial Genetics and Biotechnology
- RNA and protein synthesis mechanisms
- Ubiquitin and proteasome pathways
- Advanced NMR Techniques and Applications
- Chemical Synthesis and Analysis
- NMR spectroscopy and applications
- Bioinformatics and Genomic Networks
- Glycosylation and Glycoproteins Research
- Metabolomics and Mass Spectrometry Studies
- Computational Drug Discovery Methods
- DNA and Nucleic Acid Chemistry
- Advanced MRI Techniques and Applications
- Lipid Membrane Structure and Behavior
- Amyloidosis: Diagnosis, Treatment, Outcomes
- Peptidase Inhibition and Analysis
- Ion channel regulation and function
- Biotin and Related Studies
- RNA Research and Splicing
- Bacteriophages and microbial interactions
- Aluminum toxicity and tolerance in plants and animals
Imperial College London
2023-2024
University of Leeds
2014-2022
Institute of Structural and Molecular Biology
2014-2022
National Institute of Diabetes and Digestive and Kidney Diseases
2019-2020
National Institutes of Health
2019-2020
National and Kapodistrian University of Athens
2014
Abstract All amyloid fibrils contain a cross-β fold. How this structure differs in formed from proteins associated with different diseases remains unclear. Here, we combine cryo-EM and MAS-NMR to determine the of an fibril vitro β 2 -microglobulin (β m), culprit protein dialysis-related amyloidosis. The is composed two identical protofilaments assembled subunits that do not share m’s native tertiary fold, but are similar β-strands. motifs other fibrils, also unique features including...
Abstract Uncontrolled self-association is a major challenge in the exploitation of proteins as therapeutics. Here we describe development structural proteomics approach to identify amino acids responsible for aberrant monoclonal antibodies and design variant with reduced aggregation increased serum persistence vivo. We show that human antibody, MEDI1912, selected against nerve growth factor binds picomolar affinity, but undergoes reversible has poor pharmacokinetic profile both rat...
Significance Oligomers formed en route to amyloid fibrils are thought be the perpetrators of toxicity in many disorders. How contribute disease, however, is less clear. Here, using β 2 -micoglobulin (β m) as a model system, we show that stability highly pH-dependent, with mild acidification enhancing formation fibril-derived nonnative oligomers disrupt membranes and alter cellular function. Enhancing fibril by incubation molecular chaperone, hsp70, or cross-linking, protects against...
J-domain chaperones are involved in the efficient handover of misfolded/partially folded proteins to Hsp70 but also function independently protect against cell death. Due their high flexibility, mechanism by which they regulate cycle and how specific substrate recognition is performed remains unknown. Here we focus on DNAJB6b, has been implicated various human diseases represents a key player protection neurodegeneration protein aggregation. Using variant that exists mainly monomeric form,...
Abstract The periplasmic chaperone SurA plays a key role in outer membrane protein (OMP) biogenesis. E. coli comprises core domain and two peptidylprolyl isomerase domains (P1 P2), but its mechanisms of client binding function have remained unclear. Here, we use chemical cross-linking, hydrogen-deuterium exchange mass spectrometry, single-molecule FRET molecular dynamics simulations to map the site(s) on interrogate conformational OMP recognition. We demonstrate that samples an array...
Highlights•Dissection of protein-protein interactions in the early stages amyloid assembly•Rare biomolecular collisions and course assembly•Interaction surfaces different outcomes assembly•Molecular description species barriers assemblySummaryIn formation, heterogeneous populations oligomeric are generated, affinity, specificity, nature which may promote, inhibit, or define assembly. Despite importance intermolecular that initiate assembly, our understanding these events remains poor. Here,...
Transient oligomers are commonly formed in the early stages of amyloid assembly. Determining structure(s) these species and defining their role(s) assembly is key to devising new routes control disease. Here, using a combination chemical kinetics, NMR spectroscopy other biophysical methods, we identify structurally characterize required for protein ΔN6, truncation variant human β
Membrane forces shift the equilibria of mechanosensitive channels enabling them to convert mechanical cues into electrical signals. Molecular tools stabilize and methods capture their highly dynamic states are lacking. Cyclodextrins can mimic tension through sequestering lipids from membranes. Here we probe conformational ensemble MscS by EPR spectroscopy, lipid environment with NMR, function electrophysiology under cyclodextrin-induced tension. We show extent activation depends on...
Abstract Correct folding of outer membrane proteins (OMPs) into the Gram-negative bacteria depends on delivery unfolded OMPs to β-barrel assembly machinery (BAM). How substrates are presented BAM remains elusive, but major OMP chaperone SurA is proposed play a key role. Here, we have used hydrogen deuterium exchange mass spectrometry (HDX-MS), crosslinking, in vitro and binding assays computational modelling show that core domain one its two PPIase domains SurA-BAM interaction required for...
The balance between protein folding and misfolding is a crucial determinant of amyloid assembly. Transient intermediates that are sparsely populated during have been identified as key players in aggregation. However, due to their ephemeral nature, structural characterization these species remains challenging. Here, using the power nonuniformly sampled NMR methods we investigate pathway amyloidogenic nonamyloidogenic variants β2-microglobulin (β2m) atomic detail. Despite via common...
Significance Some of the most potent chaperones that protect against protein aggregation form heterogeneous oligomers are crucial for function. Due to polydisperse nature these oligomers, molecular origins drive self-assembly remain poorly understood. Here, using a predominantly monomeric construct C-terminal domain DNAJB6b we reveal an interplay between two N-terminal β strand configurations which analyze quantitatively in structural and kinetic terms. We show backbone conformations impact...
Protein-protein interactions (PPIs) are involved in many of life's essential biological functions yet also an underlying cause several human diseases, including amyloidosis. The modulation PPIs presents opportunities to gain mechanistic insights into amyloid assembly, particularly through the use methods which can trap specific intermediates for detailed study. Such information provide a starting point drug discovery. Here, we demonstrate that covalently tethered small molecule fragments be...
Abstract Coevolution between protein residues is normally interpreted as direct contact. However, the evolutionary record of a sequence contains rich information that may include long‐range functional couplings, couplings report on homo‐oligomeric states or even conformational changes. Due to complexity space and lack structural various members family, it has been difficult effectively mine additional encoded in multiple alignment (MSA). Here, taking advantage recent release AlphaFold (AF)...
J-domain proteins (JDPs) act as major regulators of the proteostasis network by driving specificity Hsp70 machine. Their important functions are mediated a low-complexity glycine-/phenylalanine-rich region (GF-linker) that links folded with substrate binding domain. Recently, we and others have shown in an autoinhibited JDP state, α-helix formed within GF blocks site on J-domain. However, role disordered GF-linker autoinhibition how latter is released, still not understood. Here, using...
Amyloid deposition of WT human β2-microglobulin (WT-hβ2m) in the joints long-term hemodialysis patients is hallmark dialysis-related amyloidosis. In vitro, WT-hβ2m does not form amyloid fibrils at physiological pH and temperature unless co-solvents or other reagents are added. Therefore, understanding how fibril formation initiated maintained joint space important for elucidating aggregation amyloidosis onset. Here, we investigated roles collagen I commonly administered anticoagulant,...
DNAJB6 is a prime example of an anti-aggregation chaperone that functions as oligomer. oligomers are dynamic and subunit exchange critical for inhibiting client protein aggregation. The T193A mutation in the C-terminal domain (CTD) reduces both self-oligomerization proteins, has recently been linked to Parkinson's disease. Here, we show by NMR, including relaxation-based methods, minimal effects on structure β-stranded CTD but increases population rate formation partially folded state....
Over the last 40 years nuclear magnetic resonance (NMR) spectroscopy has established itself as one of most versatile techniques for characterization biomolecules, especially proteins. Given molecular size limitations NMR together with recent advances in cryo-electron microscopy and artificial intelligence-assisted protein structure prediction, bright future structural biology been put into question. In this mini review we argue contrary. We discuss unique opportunities solution offers to...
ABSTRACT Egg envelopes of vertebrates are composed a family proteins called zona pellucida (ZP) proteins, which distinguished by the presence common structural polymerizing motif, known as ZP domain. Teleostean fish chorion is fibrous structure, consisting protein members ZPB/ZP1 and ZPC/ZP3 families, incorporated tandemly repeating heterodimers inside fibers. Computational analysis multiple from several teleostean species, reveals two potential “aggregation‐prone” sequence segments, forming...
Abstract Optimized NMR experiments are developed for isolating magnetization belonging to the I =1/2 manifolds of 13 CH 3 methyl groups in proteins, enabling manipulation a moiety as if it were an AX ( 1 H‐ C) spin‐system. These result same ‘simplification’ spin‐system that would be obtained from production { CHD 2 }‐methyl‐labeled protein samples. The sensitivity manifold‐selection is factor approximately less than corresponding acquired on }‐labeled groups. methodology described here...