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 “"Nanodisc"”.
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Structural Studies of Lipid-Bound Apolipoprotein A-I and Nanodisc-Embedded Cyp3A4 by SSNMR
… presented here focused on development of the Nanodisc platform for structural studies of membrane proteins using solid-state NMR (SSNMR).
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Multidimensional Solid-State NMR Studies of Membrane Proteins
… to systems including the membrane protein DsbB, Nanodisc, a self-assembled lipid bilayer system, and Nanodisc-embedded human cytochrome P450 3A4. For DsbB, nearly complete 13C and 15N chemical shift assignment was achieved for the transmembrane region of the protein from 3D and 4D chemical shift …
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Analysis of membrane systems using lipoprotein nanodiscs
… challenging. Engineered nanoscale lipoprotein Nanodiscs provide a well-defined and native-like lipid bilayer to solubilize membrane proteins and lipids. This dissertation details the analysis of Nanodiscs containing various membrane proteins and lipids with bioanalytical methods, including mass …
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Incorporation and analysis of bilayer constituents in nanodiscs
Nanodiscs, engineered lipoparticles composed of a lipid bilayer and two surrounding amphipathic proteins, have been utilized to house and solubilize integral membrane proteins and other molecules with large hydrophobic groups which limit their aqueous solubility. In the experiments described …
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Structure-function studies of respiratory complex I from Paracoccus denitrificans using membrane mimetics
… cryo-electron microscopy (cryo-EM). First, a nanodisc system was developed to provide the wild-type P. denitrificans complex I with a stable, membrane environment. Following optimisation of the reconstitution conditions, biochemical and biophysical characterisations confirmed the enhanced …
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Characterization of Monomeric Human Cytochrome P450 3A4 and Cytochrome P450 Reductase in Nanoscale Phospholipid Bilayer Discs
… differences between these systems of study. This Nanodisc technology allows for a homogeneous population of enzyme that is devoid of conformational heterogeneity to describe the binding and catalytic parameters without the use of detergents, whose presence can further complicate these …
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Multiplex analysis of protein-lipid and protein-membrane protein interactions utilizing silicon photonic microring resonators
… blood coagulation cascade. To accomplish this, nanodiscs were integrated with silicon photonic microring resonators. Nanodiscs are easy to assemble, nanometer scale lipid bilayer discs which offer a high degree of control over lipid and membrane composition. Each nanodisc is held together by two …
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Dynamics of protein-membrane interactions in drug metabolism and cancer
… interactions properly. Phospholipid Nanodiscs provide an excellent solution to this problem, and they allow one to control protein and lipid stoichiometry with precision. In this dissertation, I will apply Nanodisc technology to explore physiologically relevant interactions between …
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Analisi elettrofisiologica delle proteine VDAC umane incorporate nei nanodischi e indagine sui residui di cisteina dell'isoforma 3 VDAC nella funzionalità mitocondriale e nel tamponamento dei ROS
… un sistema di espressione cell-free associato a nanodisc. Questo nuovo metodo di espressione rappresenta una potente alternativa per evitare le procedure di ripiegamento in vitro delle proteine. In questo lavoro, il sistema di sintesi proteica in vitro è stato usato per la prima volta per …
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Expanding the toolbox for precision medicine with silicon photonic microring resonators and microfluidic technologies
… I developed a microfluidic platform for Nanodisc assembly and purification, referred to as the μNAP platform and detail in Chapter 5. The μNAP platform capitalizes on sample preservation and small volume processing inherent to miniaturization and microfluidics and achieves Nanodisc …
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Biochemical characterisation of the translocation of material through the protective antigen (PA) Pore
… of deuterated LFn-GFP with the both a lipid-nanodisc and a lipid-nancomplex (NC – a nanodisc containing the protective antigen pore) was followed in real time at pH7.4 and 37oC mimicking physiological conditions and at pH 5.5 mimicking the acidity of late endosomes. Measurements were …
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NON-FOULING AFFINITY PLATFORMS FOR PROTEIN IMMOBILIZATION IN ELECTRON MICROSCOPY
… GroEL from crude lysates, as well as purified nanodisc-solubilized his-malFGK2, on these NTA-modified grids with an exceptionally low level of adsorption by non-target proteins has been observed. Our data illustrates the utility of these grids for selective capture from complex mixtures, …
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Mechanistic investigation of human cytochromes P450 involved in hormone biosynthesis
… aspects of both these enzymes, I used Nanodisc technology for reconstituting active forms of these proteins in functional complexes with their effector proteins in a native-like membrane environment. Cytochrome P450 17A1 (CYP17A1) is a multi-functional enzyme, catalyzing synthesis of …
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Biophysical characterization of the Escherichia coli RNA degradosome
… in vitro studies, efforts were made to generate nanodisc supports using amphipathic copolymers. Furthermore, various mutants in the membrane-tethering segment of RNase E were created in order to stabilize the complex by crosslinking onto the lipid membrane. Using biophysical methods, the …
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Cytochrome P450 2J2 mediated metabolism of omega-6 and omega-3 endocannabinoids
… into the native membrane bilayers of nanodiscs for rigorous spectroscopic and functional studies. Notably, all N-terminal constructs were stably bound within the nanodisc bilayers, which prompted efforts to elucidate additional motifs that impart membrane binding. These observations …
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Structural insights into the GABAA receptor gating cycle
… GABA<sub>A</sub> receptors by testing multiple nanodisc scaffolds and lipid compositions. Subsequently, with the optimal condition established, I proceeded to investigate a series of GABA<sub>A</sub> receptor structures bound to β-lactam antibiotics that are postulated to act as open channel …
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Spectroscopic characterization of iron-oxygen intermediates in human aromatase (CYP19A1)
… piece together a reaction mechanism. I used the Nanodisc technology to stabilize CYP19A1 in its native membrane-like environment to obtain a mono-disperse, stable and homogenous enzyme preparation that is amicable to the optical, resonance Raman (rR) and electron paramagnetic resonance (EPR) …
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Understanding the mechanism of androgen biosynthesis
Cytochrome P45017A1 (CYP17A1) is a multifunctional steroidogenic enzyme responsible for the 17-hydroxylation of pregnenolone and progesterone as well as the subsequent 17,20 carbon-carbon bond scission of its hydroxylated products in a reaction that constitutes the first committed step of androgen …
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Synthesis and performance of novel supramolecular tools for single-particle cryogenic electron microscopy and drug and gene delivery
… from cell lysates, purified His8-GFPuv, and nanodisc embedded His6-MalFGK2. Graphene-oxide-NTA-based affinity grids were fabricated via covalent modification of graphene-oxide and monolayer fabrication on a Langmuir trough. These monolayers were characterized by AFM, SEM, and evaluated for …
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The membrane lipid composition regulates talin mediated integrin activation
… with bilayers of controlled composition using Nanodiscs technology. Results show that recruitment of talin head domain (THD) to the membrane surface is governed by electrostatics in the absence of other adapter proteins. In addition, distance measurements reveal that anionic lipids stimulate a …
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