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Showing 1 to 20 of 199 for “"cryo-em"”.
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Cryo-EM Studies of Bacterial RNA Polymerase
… The overall structure of bacterial RNAP resembles a crab claw with pincers comprising the β' and β subunits and a large cleft where the active site sits. Structural information about this essential enzyme has mainly been provided by X-ray crystal structures of stable transcription complexes. …
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Machine learning for cryo-EM structure determination
Electron cryo-microscopy (cryo-EM) has revolutionised structural biology thanks to its relaxed sample requirements. Recent advances in cryo-EM image processing have enabled the structural determination of many macromolecules that were previously intractable by other methods. At the same time, …
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Deep Learning for Segmentation Of 3D Cryo-EM Images
<p>Cryo-electron microscopy (cryo-EM) is an emerging biophysical technique for structural determination of protein complexes. However, accurate detection of secondary structures is still challenging when cryo-EM density maps are at medium resolutions (5-10 Å). Most existing methods are image …
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2D image classification and alignment in single-particle Cryo-EM
Background and Objective: Single-particle Cryo-EM is a powerful tool for structural biologists to determine structures of macro-molecules at atomic resolution. In single-particle Cryo-EM, many individual biological molecular particles with identical structures are rapidly frozen at unknown, random …
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A Geometric Approach for Deciphering Protein Structure from Cryo-EM Volumes
Electron Cryo-Microscopy or cryo-EM is an area that has received much attention in the recent past. Compared to the traditional methods of X-Ray Crystallography and NMR Spectroscopy, cryo-EM can be used to image much larger complexes, in many different conformations, and under a wide range of …
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Machine Learning for Reconstructing Dynamic Protein Structures from Cryo-EM Images
… of dynamic biomolecular complexes via cryo-electron microscopy (cryo-EM). In single particle cryo-EM, the central problem is to reconstruct the 3D structure of a target biomolecular complex from a set of noisy and randomly oriented 2D projection images, a challenging inverse problem …
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A Graph-Based Algorithm to Determine Protein Structure from Cryo-EM Data
Cryo-electron microscopy: cryo-EM) provides 3D density maps of proteins, but these maps do not have sufficiently high resolution to directly yield atomic-scale models. Previous work has shown that features known as secondary structures can be located in these density maps. A second source of …
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Cryo-EM studies of substrate and inhibitor binding to mammalian respiratory complex I
… an intricate multi-subunit, energy-transducing membrane protein that is essential for aerobic energy metabolism and NADH/NAD⁺ homeostasis. It couples the energy released from NADH oxidation and ubiquinone (Q) reduction to pump four protons across the inner mitochondrial membrane, contributing to …
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Methods for time-resolved cryo-EM using pulsed infrared radiation and vapour phase deposition
Electron cryo-microscopy (cryo-EM) is an imaging technique for determining high spatial resolution structures of proteins. Technological advances have made it possible to attain atomic resolution and attention may now be directed towards the challenge of extending the time resolution of cryo-EM …
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The 2.7 resolution cryo-EM reconstruction of Mycobacterium tuberculosis encapsulin nanocompartment containing DyP peroxidase
… about the structure and function of this system. Both encapsulin and DyP were expressed and purified recombinantly in E. coli. Cryo- EM particle processing and EM map reconstruction was carried in an attempt to generate a density map of encapsulated DyP along with model building. Native …
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DNA-PKcs/ Artemis Complex in DNA Double-Strand-Break Repair: Cryo-EM and Biochemical Studies
… by the cell cycle as it does not require a template for recombination. Also, NHEJ has been shown to be the preferred DSB repair pathway in higher eukaryotic organisms including human. NHEJ is dynamic and flexible but can be separated into three steps - DNA end recognition, end synapsis and …
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Cryo-EM structural exploration of human VPS34 complexes and their activation by RAB GTPases and NRBF2
… into phosphatidylinositol-3-phosphate (PI3P) on membranes. The reaction product, PI3P, is a potent signalling lipid involved promoting autophagy, phagocytosis, and endocytic sorting in mammalian cells. In addition to its roles in normal homeostasis, VPS34 can be hijacked by viruses and bacteria …
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Cryo-EM studies of class A GPCRs: the thyroid-stimulating hormone receptor and the β1-adrenoceptor
G protein-coupled receptors (GPCRs) are integral membrane proteins with seven transmembrane domains. They are essential in intercellular communication in human physiology and represent important drug targets in human disease. While methodological advances, particularly in cryogenic electron …
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Cryo-EM reveals how adeno-associated virus engages its receptor AAVR and an investigation of the GPR108 ectodomain
… AAV engages with human cells and the immune system we hope to create a roadmap of the AAV capsid that can help direct AAV capsid mutations for a more efficient AAV capsid, which can more easily evade the immune system, while maintaining its essential cellular interactions. We have previously …
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Cryo-EM as a Tool to Study the Diverse Interactome of Ku 70/80 in NHEJ and Beyond
… of damage, DSBs are the most perilous (3). If remained unrepaired or repaired incorrectly, they can be drivers for chromosomal rearrangements, cellular senescence, and apoptosis (2, 4). Despite its fundamental role in genomic stability, NHEJ has been implicated in carcinogenesis, tumour …
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Affinity cryo-electron microscopy: Methods development and applications
<p>Single particle cryo-electron microscopy (cryo-EM) is an emerging powerful tool for structural studies of macromolecular assemblies. Although less concentrated and smaller amounts of samples are required for single particle cryo-EM compared to X-ray crystallography, it remains challenging to …
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Computational Development for Secondary Structure Detection From Three-Dimensional Images of Cryo-Electron Microscopy
<p>Electron cryo-microscopy (cryo-EM) as a cutting edge technology has carved a niche for itself in the study of large-scale protein complex. Although the protein backbone of complexes cannot be derived directly from the medium resolution (5-10 Å) of amino acids from three-dimensional (3D) density …
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INVESTIGATION OF CRYOGENIC SURFACE CHARGING DURING OFF-AXIS ELECTRON HOLOGRAPHY
Cryogenic electron microscopy (cryo-EM) is an increasingly popular technique for determining the structure of solutions, especially of those with protein solutes. The ability to immobilize a fluid and attain atomic resolution of solutes within has propelled the technique to the forefront of …
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Synthesis and performance of novel supramolecular tools for single-particle cryogenic electron microscopy and drug and gene delivery
… drug design and basic research into complex biochemical processes. Cryo-EM is an emerging alternative to XRD and NMR that is complementary in many ways relative to XRD and NMR. Materials approaches to cryo-EM are anticipated to greatly facilitate the cryo-EM process, allowing progress toward a …
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Using molecular approaches to understand Complex I deficiency in mouse models
… proton pumping across the mitochondrial inner membrane, and generates the proton motive force that drives ATP synthesis and transport processes. The ~1 MDa mammalian complex contains 45 subunits, and pathological mutations in both its mitochondrial and nuclear encoded subunits result in diverse …
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