A5090746349- Photosynthetic Processes and Mechanisms
- ATP Synthase and ATPases Research
- Mitochondrial Function and Pathology
- Metalloenzymes and iron-sulfur proteins
- Electrocatalysts for Energy Conversion
- Bacteriophages and microbial interactions
- RNA and protein synthesis mechanisms
- Bacterial Genetics and Biotechnology
- Glycosylation and Glycoproteins Research
- Microbial Community Ecology and Physiology
- Photoreceptor and optogenetics research
- Catalysis for Biomass Conversion
- RNA modifications and cancer
- Genomics and Phylogenetic Studies
- Carbohydrate Chemistry and Synthesis
- Advanced Proteomics Techniques and Applications
- Adipose Tissue and Metabolism
- Antibiotic Resistance in Bacteria
- Analytical Chemistry and Chromatography
- Amino Acid Enzymes and Metabolism
- Escherichia coli research studies
- Environmental Chemistry and Analysis
- Advanced battery technologies research
- Protein Tyrosine Phosphatases
- Microbial Metabolic Engineering and Bioproduction
University of York
2019-2025
MRC Mitochondrial Biology Unit
2012-2023
University of Cambridge
2017-2023
Biomedical Research Institute
2022
ORCID
2021
Medical Research Council
2012-2020
Wellcome Trust
2015-2018
Google (United States)
2018
Bacterial efflux pumps confer multidrug resistance by transporting diverse antibiotics from the cell. In Gram-negative bacteria, some of these form multi-protein assemblies that span cell envelope. Here, we report near-atomic resolution cryoEM structures Escherichia coli AcrAB-TolC pump in resting and drug transport states, revealing a quaternary structural switch allosterically couples synchronizes initial ligand binding with channel opening. Within transport-activated state, remains open...
The molecular mode of action biguanides, including the drug metformin, which is widely used in treatment diabetes, incompletely characterized. Here, we define inhibitory drug-target interaction(s) a model biguanide with mammalian respiratory complex I by combining cryo–electron microscopy and enzyme kinetics. We interpret these data to explain selectivity binding different states. primary site an amphipathic region quinone-binding channel, additional pocket on intermembrane-space side...
The mitochondrial genome (mtDNA) encodes essential machinery for oxidative phosphorylation and metabolic homeostasis. Tumor mtDNA is among the most somatically mutated regions of cancer genome, but whether these mutations impact tumor biology debated. We engineered truncating mtDNA-encoded complex I gene, Mt-Nd5, into several murine models melanoma. These promoted a Warburg-like shift that reshaped microenvironments in both mice humans, consistently eliciting an anti-tumor immune response...
Abstract To conserve energy during starvation and stress, many organisms use hibernation factor proteins to inhibit protein synthesis protect their ribosomes from damage 1,2 . In bacteria, two families of factors have been described, but the low conservation these huge diversity species, habitats environmental stressors confounded discovery 3–6 Here, by combining cryogenic electron microscopy, genetics biochemistry, we identify Balon, a new in cold-adapted bacterium Psychrobacter urativorans...
Significance Mitochondria produce ATP by using respiration to drive synthase. Respiration is catalyzed several membrane-bound complexes that are structurally organized into supercomplex assemblies. Supercomplexes have been proposed confer a catalytic advantage channeling of substrates between enzymes in the Here, we test three simple predictions behavior mammalian respiratory chain depend on whether supercomplexes kinetically important and show it not. We reinterpret previous data taken...
Highlights•Preparation of mammalian complex I in the deactive state that forms during ischemia•The structure determined using electron cryomicroscopy•Improved particle densities and orientations obtained PEGylated gold grids•Localized unfolding around quinone-binding site stateSummaryComplex (NADH:ubiquinone oxidoreductase) is central to energy metabolism mitochondria. It couples NADH oxidation by ubiquinone proton transport across energy-conserving inner membrane, catalyzing respiration...
Abstract Respiratory complex I (NADH:ubiquinone oxidoreductase) captures the free energy from oxidising NADH and reducing ubiquinone to drive protons across mitochondrial inner membrane power oxidative phosphorylation. Recent cryo-EM analyses have produced near-complete models of mammalian complex, but leave molecular principles its long-range coupling mechanism open debate. Here, we describe 3.0-Å resolution structure mouse heart mitochondria with a substrate-like inhibitor, piericidin A,...
Abstract Type II NADH:quinone oxidoreductase (NDH-2) is central to the respiratory chains of many organisms. It not present in mammals so may be exploited as an antimicrobial drug target or used a substitute for dysfunctional complex I neuromuscular disorders. NDH-2 single-subunit monotopic membrane protein with just flavin cofactor, yet no consensus exists on its mechanism. Here, we use steady-state and pre-steady-state kinetics combined mutagenesis structural studies determine mechanism...
Electron bifurcation is a fundamental energy conservation mechanism in nature which two electrons from an intermediate-potential electron donor are split so that one sent along high-potential pathway to acceptor and the other low-potential acceptor. This process allows endergonic reactions be driven by exergonic ones alternative, less recognized, of coupling well-known chemiosmotic principle. The electron-bifurcating [FeFe] hydrogenase Thermotoga maritima (HydABC) requires both NADH...
Abstract CO 2 fixation is commonly limited by inefficiency of the -fixing enzyme Rubisco. Eukaryotic algae concentrate and fix in phase-separated condensates called pyrenoids, which complete up to one-third global fixation. Condensation Rubisco pyrenoids dependent on interaction with disordered linker proteins that show little conservation between species. We developed a sequence-independent bioinformatic pipeline identify green algae. report from Chlorella demonstrate it binds conserved...
Mitochondrial complex I (NADH:ubiquinone oxidoreductase), a crucial enzyme in energy metabolism, captures the redox potential from NADH oxidation/ubiquinone reduction to create proton motive force used drive ATP synthesis oxidative phosphorylation. High-resolution single-particle electron cryo-EM analyses have provided detailed structural knowledge of catalytic machinery I, but not molecular principles its transduction mechanism. Although ubiquinone is considered bind long channel at...
Abstract Complex I is a crucial respiratory enzyme that conserves the energy from NADH oxidation by ubiquinone‐10 (Q 10 ) in proton transport across membrane. Studies of its transduction mechanism are hindered extreme hydrophobicity Q , and they have so far relied on native membranes with many components or hydrophilic analogues partition into undergo side reactions. Herein, we present self‐assembled system without these limitations: proteoliposomes containing mammalian complex I, quinol...
Abstract The O-linked β-N-acetylglucosamine modification is a core signalling mechanism, with erroneous patterns leading to cancer and neurodegeneration. Although thousands of proteins are subject this modification, only single essential glycosyltransferase catalyses its installation, the O-GlcNAc transferase, OGT. Previous studies have provided truncated structures OGT through X-ray crystallography, but full-length protein has never been observed. Here, we report 5.3 Å cryo-EM model We show...
Abstract CO 2 fixation is commonly limited by inefficiency of the -fixing enzyme Rubisco. Eukaryotic algae concentrate and fix in phase-separated condensates called pyrenoids, which complete up to one-third global fixation. Condensation Rubisco pyrenoids dependent on interaction with disordered linker proteins that show little conservation between species. We developed a sequence-independent bioinformatic pipeline identify green algae. report from Chlorella demonstrate it binds conserved...
Bacterial spore germination transforms spores from dormant bodies to vegetative cells. Initiation of or rehydration can proceed without exogenous energy sources, so bioenergetic processes in have been overlooked, despite many germinants being such as sugars. Here, we apply remission spectroscopy noninvasively measure the energy-transducing electron-transport chain intact during germination. In Bacillus megaterium and subtilis, find energisation cytoplasmic metabolism electron transport...
ABSTRACT Microviruses are single-stranded DNA viruses infecting bacteria, characterized by T = 1 shells made of single jelly-roll capsid proteins. To understand how microviruses infect their host cells, we have isolated and studied an unusually large microvirus, Ebor. Ebor belongs to the proposed “Tainavirinae” subfamily Microviridae infects model Alphaproteobacterium Rhodobacter capsulatus . Using cryogenic electron microscopy, show that enlarged is result extended C-terminus major protein....
Trypanosoma brucei is the causal agent of African trypanosomiasis in humans and animals, latter resulting significant negative economic impacts afflicted areas world. Resistance has arisen to trypanocidal drugs pentamidine melarsoprol through mutations aquaglyceroporin TbAQP2 that prevent their uptake. Here we use cryogenic electron microscopy determine structure from , bound either substrate glycerol or sleeping sickness drugs, melarsoprol. The bind within AQP2 channel at a site completely...
Metal-dependent formate dehydrogenases (FDHs) catalyze the reversible conversion of into CO2, a proton, and two electrons. Kinetic studies FDHs provide key insights their mechanism catalysis, relevant as guide for development efficient electrocatalysts oxidation well CO2 capture utilization. Here, we identify explain kinetic isotope effect (KIE) observed deuterioformate by Mo-containing FDH from Escherichia coli using three different techniques: steady-state solution assays, protein film...
In oxidative phosphorylation, ATP synthases interconvert two forms of free energy: they are driven by the proton-motive force across an energy-transducing membrane to synthesize and displace ADP/ATP ratio from equilibrium. For thermodynamically efficient energy conversion must be reversible catalysts. However, in many species unidirectional catalysts (their rates hydrolysis negligible), others mechanisms have evolved regulate or minimize hydrolysis. Unidirectional catalysis Paracoccus...