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Showing 1 to 20 of 135 for “"cryo-electron microscopy"”.
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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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Cryo-electron microscopy studies of pluripotency factor Nanog
… oligomerisation of Nanog drives self-renewal, cryo-electron microscopy and single particle analysis were utilised to determine a 3D structure of Nanog. By performing size exclusion chromatography and analytical ultracentrifugation, it was discovered that Nanog forms large oligomers of 860 kDa …
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Cryo-electron Microscopy of Reconstituted Membrane Fusion Systems
… of a SNARE reconstituted in-vitro system, I use cryo-electron tomography to provide insight into how SNARE proteins organise at fusion sites showing that such organisation occurs in a vesicle-plasma membrane distance-dependent manner. In order to improve the quality of structural data acquired at …
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Cryo-electron microscopy studies on ovine mitochondrial complex I
… oxidative phosphorylation pathway. It couples electron transfer between NADH and ubiquinone to proton translocation across the inner mitochondrial membrane, contributing to cellular energy production. Complex I is the largest and most elaborate protein assembly of the respiratory chain with a …
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Cryo-electron microscopy image analysis using multi-frequency vector diffusion maps
Cryo-electron microscopy (EM) single particle reconstruction is an entirely general technique for 3D structure determination of macromolecular complexes. However, because the images are taken at low electron dose, it is extremely hard to visualize the individual particle with low contrast and high …
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Understanding the Structure and Function of GABAA Receptors Using Cryo-Electron Microscopy
The general metadata -- e.g., title, author, abstract, subject headings, etc. -- is publicly available, but access to the submitted files is restricted to UT Southwestern campus access and/or authorized UT Southwestern users.
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Advances In Computer Aided Protein Structure Determination From Sparse Cryo Electron Microscopy Data
Single-particle cryo-electron microscopy (cryoEM) has become a powerful tool for determining macromolecular structures. Thanks to recent advances in direct electron detectors and motion correction algorithms it can frequently deliver electron density maps in the range of 3-5Å resolution. To obtain …
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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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Cryo-electron microscopy of HPV16 pseudovirions reveal changes in capsid conformation upon furin cleavage
… from cleavage of L2 by exogenous furin using cryoelectron microscopy (cryo-EM). At 5 Å resolution, we observed that furin-cleaved HPV16 PsVs capsids display widespread changes in the arrangement of capsomeres relative to uncleaved control virions. This structural change is relevant because …
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Segmentation and registration of molecular components in 3-dimensional density maps from cryo-electron microscopy
Cryo-electron microscopy is a method that produces 3D density maps of macromolecular complexes. Segmentation and registration methods are heavily used to extract structural information from such density maps. Segmentation aims to identify regions in a density map corresponding to individual …
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Multi-Modal Regualtion of Actin Networks
… simultaneous optical trapping and confocal microscopy assays and in vitro reconstituted myosin motor-based total internal reflection fluorescence microscopy assays were employed to study how mechanical forces applied on actin filaments can regulate actin binding. A case study on a homologous …
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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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Unraveling the Functions of Synaptotagmin and Munc13 in Neurotransmitter Release
… of membrane bridging by synaptotagmin. Our cryo-electron microscopy (cryo-EM) images showed that a majority of synaptotagmin fragment containing both C¬2A and C2B domains (C2AB) molecules bridge membranes directly. Fluorescence spectroscopy demonstrates that the bottom of the C2B domain …
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Structures, Functions, and Druggability of the SARS-CoV-2 Replication–Transcription Complex
… substrate (referred to as the nsp132-RTC) using cryo-electron microscopy (cryo-EM). Intriguingly, we observed that one of the bound helicases is orientated with an opposing polarity to the RdRp on the template RNA (t-RNA) strand. In chapter III, we investigated the effect of this configuration on …
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Structual Visualization of Cytoskeletal Force Tranduction
… protein α-catenin.In vitro reconstitution and cryo-electron microscopy reveal myosin force-evoked superhelical F-actin spirals. Three-dimensional reconstruction and variability analysis uncover extensive asymmetric remodeling of F-actin's helical lattice. This is recognized by α-catenin, which …
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Lengthening the timescale reach of molecular dynamics
… experimental data from crystallography and cryo-electron microscopy to explore the functional dynamics of biological systems. The timescale reach of the molecular dynamics tool is limited by how fast femtosecond timesteps can be sequentially integrated; today's fast computers allow …
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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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Structural Investigation of the Chromatin Remodeller Fun30
… structural characterisation especially using cryo electron microscopy.
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Structural analyses of the ribosome by hybrid approach
… experimental data from X-ray crystallography and cryo-electron microscopy with computing, permitting one to resolve atomic-level structures of intermediates of the functional ribosome and, thereby, to advance our understanding of ribosome function and the underlying physical mechanisms. In this …
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