A5039569974- Protein Structure and Dynamics
- Lipid Membrane Structure and Behavior
- Alzheimer's disease research and treatments
- Metabolomics and Mass Spectrometry Studies
- Enzyme Structure and Function
- RNA and protein synthesis mechanisms
- Bacterial Genetics and Biotechnology
- Fibroblast Growth Factor Research
- Chemical Reaction Mechanisms
- Photosynthetic Processes and Mechanisms
- Advanced NMR Techniques and Applications
- Cardiomyopathy and Myosin Studies
- Advanced ceramic materials synthesis
- Computational Drug Discovery Methods
- Chemical Synthesis and Analysis
- Bacteriophages and microbial interactions
- Porphyrin Metabolism and Disorders
- Molecular spectroscopy and chirality
- Prion Diseases and Protein Misfolding
- Diet and metabolism studies
- DNA and Nucleic Acid Chemistry
- Monoclonal and Polyclonal Antibodies Research
- Mass Spectrometry Techniques and Applications
- Parasitic Infections and Diagnostics
- Microtubule and mitosis dynamics
University of Kent
2019-2025
Wellcome Trust
2020-2025
University of Leeds
2008-2019
Institute of Structural and Molecular Biology
2002-2019
Lehigh University
1996-2016
Universität Innsbruck
2014
Université Grenoble Alpes
2014
International AIDS Society
2014
Institut de Biologie Structurale
2014
Université Laval
2014
Numerous studies of amyloid assembly have indicated that partially folded protein species are responsible for initiating aggregation. Despite their importance, the structural and dynamic features amyloidogenic intermediates molecular details how they cause aggregation remain elusive. Here, we use ΔN6, a truncation variant naturally β2-microglobulin (β2m), to determine solution structure nonnative intermediate at high resolution. The ΔN6 reveals major repacking hydrophobic core accommodate...
Abstract Protein tyrosine kinases differ widely in their propensity to undergo rearrangements of the N-terminal Asp–Phe–Gly (DFG) motif activation loop, with some, including FGFR1 kinase, appearing refractory this so-called ‘DFG flip’. Recent inhibitor-bound structures have unexpectedly revealed for first time a ‘DFG-out’ state. Here we use conformationally selective inhibitors as chemical probes interrogation structural and dynamic features that appear govern DFG flip FGFR1. Our detailed...
Using a combination of experimental techniques (circular dichroism, differential scanning calorimetry, and NMR) molecular dynamics simulations, we performed an extensive study denaturation the Trp-cage miniprotein by urea guanidinium. The experiments, despite their different sensitivities to various aspects process, consistently point simple, two-state unfolding process. Microsecond simulations with femtosecond time resolution allow us unravel detailed mechanism unfolding. process starts...
Current interferon-based therapy for hepatitis C virus (HCV) infection is inadequate, prompting a shift toward combinations of direct-acting antivirals (DAA) with the first protease-targeted drugs licensed in 2012. Many compounds are pipeline yet primarily target only three viral proteins, namely, NS3/4A protease, NS5B polymerase, and NS5A. With concerns growing over resistance, broadening repertoire DAA targets major priority. Here we describe complete structure HCV p7 protein as monomeric...
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,...
Nuclear magnetic resonance (NMR) is a powerful tool for observing the motion of biomolecules at atomic level. One technique, analysis relaxation dispersion phenomenon, highly suited studying kinetics and thermodynamics biological processes. Built on top relax computational environment NMR dynamics new designed to be comprehensive, accurate easy-to-use. The software supports more models, both numeric analytic, than current solutions. An automated protocol, available scripting driving...
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
Chloride Intracellular Channel 1 (CLIC1) is a metamorphic protein capable of transitioning from soluble cytoplasmic state to membrane-bound chloride channel. This conformational shift, crucial for physiological processes such as cell volume regulation, electrical excitability, and angiogenesis, linked pathological conditions including malignancies cardiovascular diseases. Despite its significance, the molecular mechanism driving CLIC1 membrane insertion has remained elusive. Using an...
Background: Blastocystis, the most prevalent microbial eukaryote in humans, has a global distribution. Studies have linked its presence with distinct gut microbiome and metabolome profiles compared to those where organism is absent. However, interplay of antibiotics administration, Blastocystis surrounding remains understudied. This case study aimed explore antibiotic consumption subsequent changes an individual diagnosed irritable bowel syndrome (IBS). Methods: Stool samples from IBS...
Background: Blastocystis, the most prevalent microbial eukaryote in humans, has a global distribution. Studies have linked its presence with distinct gut microbiome and metabolome profiles compared to those where organism is absent. However, interplay of antibiotics administration, Blastocystis surrounding remains understudied. This case study aimed explore antibiotic consumption subsequent changes an individual diagnosed irritable bowel syndrome (IBS). Methods: Stool samples from IBS...
Resistance to the antibiotic fusidic acid (FA) in human pathogen Staphylococcus aureus usually results from expression of FusB-type proteins (FusB or FusC). These bind elongation factor G (EF-G), target FA, and rescue translation FA-mediated inhibition by an unknown mechanism. Here we show that FusB family are two-domain metalloproteins, C-terminal domain which contains a four-cysteine zinc finger with unique structural fold. This mediates high-affinity interaction domains EF-G. By binding...
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...
Coenzyme A (CoA) is an ubiquitous and essential cofactor, synthesized from the precursor pantothenate. Vitamin biosynthetic pathways are normally tightly regulated, including pathway pantothenate to CoA. However, no regulation of biosynthesis has been identified. We have recently described additional component in pathway, PanZ, which promotes activation zymogen, PanD, form aspartate α-decarboxylase (ADC) a CoA-dependent manner. Here we report structure PanZ complex with reveals structural...
The unfolded ensemble in aqueous solution represents the starting point of protein folding. Characterisation this species is often difficult since native state usually predominantly populated at equilibrium. Previous work has shown that four-helix protein, Im7 (immunity 7), folds via an on-pathway intermediate. While transition states and folding intermediate have been characterised atomistic detail, knowledge under same ambient conditions remained sparse. Here, we introduce destabilising...
Abstract Antibiotic resistance in clinically important bacteria can be mediated by proteins that physically associate with the drug target and act to protect it from inhibitory effects of an antibiotic. We present here first detailed structural characterization such a protection mechanism through protein-protein interaction, revealing architecture complex formed between FusB fusidic acid protein (EF-G) acts protect. Binding EF-G induces conformational dynamic changes latter, shedding light...
We present a simple methodology to assess the phospholipid selectivity of membrane-associated compounds in natural membranes.
By use of NMR residual dipolar coupling measurements in a dilute liquid-crystalline solvent, the solution structure has been determined complex between oligosaccharide moiety globotriaosylceramide (Gb(3)-OS) and B-subunit homopentamer verotoxin 1 (VTB). The data indicate that Gb(3)-OS binds single binding site per monomer, which is identical to one three sites inferred from X-ray same complex. We find no evidence within experimental error for occupancy at either two additional observed...
In studies on the thermodynamics of ligand-protein interactions, it is often assumed that configurational and conformational entropy ligand zero in bound state (i.e., rigidly fixed binding pocket). However, there little direct experimental evidence for this assumption, case p-substituted benzenesulfonamide inhibitors to bovine carbonic anhydrase II (BCA II), observed thermodynamic signature derived from isothermal titration calorimetry experiments leads indirectly conclusion a considerable...
Quantifying phospholipid bilayer-small molecule interactions is vital to the development of new drug candidates and/or medicinal therapies. However, obtaining these data remains problematic. Herein, we detail a nanodisc assay which enables elucidation using conventional solution state NMR spectroscopy techniques.
Abstract Background Blastocystis , the most prevalent eukaryotic gut microbe in humans, has a global distribution. Studies have linked its presence with distinct microbiome and metabolome profiles compared to those where organism is absent. However, vivo effect of antibiotics on surrounding remains understudied. This case study aimed explore how antibiotic consumption influences subsequent changes an individual irritable bowel syndrome (IBS). Methods Stool samples from IBS patient, collected...