Global ETD Search
Search theses and dissertations gathered from participating repositories worldwide. Every result links back to the library that holds it. No account is needed.
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Showing 1 to 20 of 24 for “"cryo-electron tomography"”.
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Investigating Microtubules Inside Cells with Cryo-Electron Tomography
… of neuronal microtubules remain unclear. Cryogenic electron tomography allows for visualization and structure determination of purified or cellular specimen in their native-like, vitrified state. I used this method in combination with subtomogram averaging to study intracellular …
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Visualisation of innate immune signalling complexes in cells by cryo-electron tomography
… labelling experiments of MAVS. I optimised a cryo-correlative light electron microscopy (cryo-CLEM) workflow to attain high-resolution structural insights into the ASC signalling complexes in cells. My cryo-ET reconstructions, obtained in unstained and fully hydrated conditions, show that the …
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Investigating the cytoskeleton and microtubule-based transport with in situ cryo-electron tomography
… of cargo interaction are not well described. Cryo-electron tomography (cryo-ET) can be applied to cellular samples to resolve macromolecules spatially at nanometer resolution. When many copies of a target are present, sub-tomogram averaging (STA) can be applied to improve resolution. I aimed …
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A CORRELATIVE CRYO-ELECTRON TOMOGRAPHY APPROACH TO STUDY BRD4-NUT CHROMATIN CONDENSATES ACROSS SCALES
… their compatibility with active transcription. Cryo-electron tomography (cryo-ET) is suitable for conducting such analyses, as it allows molecular-resolution imaging of cellular structures within their native context. However, methodological challenges limit the straightforward application of …
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Structural and biochemical characterization of nuclear pore complex structural scaffold sub-complexes
… of the overall architecture of the NPC by cryo-electron tomography(cryo-ET) to ~23Å resolution has revealed its size, shape, and course arrangement, but lacks distinguishable protein-protein boundaries and secondary structural details.
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Visualisation of dynein complexes in vitro and inside cells
… is also not clear. To understand this, I used cryo-electron microscopy (EM) combined with biochemical analysis and fluorescence microscopy to study dynein in vitro and in cells. In Chapter 2, I present the cryo-EM structure of purified full-length human cytoplasmic-1 in an inhibited state. By …
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Computational Modeling of Claudin-15 Paracellular Channels
… simulations, reduced computational modeling, and cryo-electron tomography (cryo-ET) to address these gaps. Atomistic MD simulations of Cldn-15 strands in opposing plasma membranes demonstrated that lateral flexibility emerges from multiple modular cis interfaces, permitting curvature and branching …
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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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Investigating the role of ZEB2 in the establishment of neuroepithelial architecture and axon tract formation
… study the ultrastructure of navigating axons by cryo-electron tomography.
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Mechanistic Insights into the Role of Munc13 in Synaptic Vesicle Docking, Priming, and Fusion
… reconstitutions, dynamic light scattering, and cryo-electron tomography I explored the functional relevance of membrane binding within the C1, C2B, and C2C domains of Munc13. The C2C domain was identified as a critical component of Munc13 that enables bridging between liposomes with synaptic …
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Developmental Changes In The Structure and Composition of The Postsynaptic Density
… are identical. Here, immuno-gold labeling and electron tomography of PSDs isolated throughout development was used to track changes in both the structure and molecular composition of the PSD. State-of-the-art cryo-electron tomography was used to study the fine structure of the PSD during …
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Molecular Mechanisms Involved in Chromosome Segregation
… yeast (known as Spindle Pole Body) and performed cryo-electron tomography analysis of native γ-TuRC complexes nucleating microtubules. We observed that γ-TuRC is a perfect symmetry match for MT nucleation and that additional factors allow efficient MT nucleation at native γ-TuRCs.
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Structural studies of SARS-CoV-2 spike protein and vesicular coats
… Lastly, in another collaboration, I used cryo-electron microscopy and single particle analysis to characterise a series of antiviral peptides and their interaction with the S protein in solution as well as on intact virions. In the second part, I discuss my work on vesicular coat proteins. …
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ADVANCING IN SITU CRYO-ET TO THE NEXT DIMENSION: TOMOGRAPHY ON 3D ORGANOIDS
… remains a central challenge in biology. In situ cryo-electron tomography (cryo-ET) can help to address this challenge by providing high-resolution, three-dimensional visualization of macromolecules in near-native, minimally disturbed conditions. With its broad resolution, cryo-ET enables us to …
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The encapsidation protein from Lactococcus lactis asccφ28 as a novel system for the study of molecular motors
… preliminary crystallographic, as well as cryo-electron tomography (cryo-ET) and cryo-electron microscopy (cryo-EM) studies, suggest the decameric assembly is in fact composed of a dimer of pentamers, akin to the widely investigated and closest related EP from phage φ29, which forms …
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Structural and Spatiotemporal Organisation of Bax and Apaf1 during Apoptosis
… of the mitochondrion architecture. Using cryo-electron tomography, Bax clusters were shown to form distinct cellular environments organised in a meshwork similar to a sponge-like structure that might be bound to the ruptured outer mitochondrial membrane. This suggests that Bax may …
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Molecular modeling and simulation of bacterial chemosensory arrays
… structural data from X-ray crystallography and cryo-electron tomography (cryo-ET), an atomic model of the cytoplasmic portion of the chemosensory array from Thermotoga maritima is constructed and refined. Through the use of large-scale MD simulations, a novel conformational change in a key …
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Structure & Function of Bacterial Transport Machines in their Cellular Context
… test purification strategies for single-particle cryo-electron microscopy studies. Finally, I apply further bioinformatic approaches and synthesise my computational and experimental results to propose a mechanism of transport and suggest future experimental tests. I conduct MD simulations of MacB …
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Fantastic ribonucleoprotein complexes and how to study them. Architectural principles of regulatory assemblies in bacteria.
… corresponding mRNA molecules. In this thesis, Cryo-EM studies on target RNAs such as amiE and rbsB, reveal how Hfq presents multiple A-rich motifs in the mRNA 5ʹ-end to Crc and triggers assembly of higher order complexes. The structures suggest that secondary structure elements and the sequence …
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Towards structural characterisation of ciliary rootlets, ciliary caps and intraflagellar transport
… in situ anterograde IFT-trains was determined by cryo-electron tomography (cryo-ET) in the Pigino lab, but high-resolution structural information of the interactions within and between the IFT-A and IFT-B subcomplexes remains absent. I set out to purify IFT-complexes from the motile cilia of the …
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