Tesis sobre el tema "Conformational characterization"
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Ambrogi, Martina. "Synthesis, characterization and conformational studies of arylbenzylmaleimides". Master's thesis, Alma Mater Studiorum - Università di Bologna, 2012. http://amslaurea.unibo.it/4251/.
Texto completoSilenzi, Ilaria. "Synthesis, characterization and conformational studies of bis-phenothiazine-aryl-boranes". Master's thesis, Alma Mater Studiorum - Università di Bologna, 2019. http://amslaurea.unibo.it/19201/.
Texto completoZych, Andrew John. "Conformational characterization of abiotic secondary structure based on aromatic stacking /". Full text (PDF) from UMI/Dissertation Abstracts International, 2001. http://wwwlib.umi.com/cr/utexas/fullcit?p3008484.
Texto completoFoschi, Simone. "Synthesis, Characterization and Conformational Studies of Modified Carbazole-bis-aryl-boranes". Master's thesis, Alma Mater Studiorum - Università di Bologna, 2020. http://amslaurea.unibo.it/20686/.
Texto completoCroasdale, Rebecca Alice. "Characterization of the conformational dynamics of the Nek2 leucine zipper domain". Thesis, University of Leicester, 2013. http://hdl.handle.net/2381/27799.
Texto completoIeronymaki, Matthaia. "Immunological and Conformational characterization of synthetic peptide probes for autoimmune diseases". Thesis, Cergy-Pontoise, 2016. http://www.theses.fr/2016CERG0831/document.
Texto completoAutoimmune diseases (ADs) refer to chronic and heterogeneous diseases with acquired immune system’s reactions against the body’s own healthy tissues. ADs affect more than 5% of the population worldwide and especially young adults. The complexity of their spectrum is enormous and even if their etiology is still unclear, it was demonstrated that both genetic and environmental factors are involved in triggering the pathological mechanism. Hence, a reliable diagnostic and/or prognostic tool for an early diagnosis of ADs before irreversible cellular damage occurs and for monitoring their progression is demanded.Numerous studies have revealed the presence of different autoantibodies (auto-Abs) in sera of patients suffering from ADs. Autoantibodies that are specific for a disease can be used as biomarkers (BMs) for its diagnosis while autoantibodies that differ depending on the disease state can be used in the follow up of the patients. Actually, in the case of autoimmunity, an easily detectable and reliable BM may be represented by the titer of a specific auto-Ab.In this context, we aimed to identify target(s) of the response for two different ADs, multiple sclerosis (MS) and monoclonal gammopathy, using the chemical reverse approach, which involves the screening of focused antigen (Ag) libraries with patients’ serum.In particular, the significance of anti-myelin antibodies, and especially, anti- Myelin Oligodendrocyte Glycoprotein (anti-MOG) antibodies is still matter of debate, underscoring the highly controversial issue of a putative pathogenetic role of anti-MOG antibodies in MS. In this thesis we investigated the role of MOG as putative auto-Ag in MS using the experimental autoimmune encephalomyelitis (EAE) model. Moreover, in order to assess the presence of a B-cell epitope spreading mechanism, i.e. the occurrence of a response directed toward epitopes distinct from the disease-inducing agent, we synthesized and tested as antigenic probes also five synthetic peptides covering the 1-117 sequence of MOG.The second issue focused on the selection of a peptide mimicking the minimal epitope recognized by the commercial available monoclonal antibody anti-human natural killer cell-1 (anti-HNK-1) using Surface Plasmon Resonance (SPR) technique. HNK-1 epitope, is considered as the antigenic determinant of myelin-associated glycoprotein (MAG), a quantitatively minor component of myelin sheaths. It is observed that patients affected by autoimmune neurological disorders, such as IgM monoclonal gammopathy and demyelinating polyneuropathy, often develop anti-MAG antibodies specifically targeting the HNK-1 epitope. Accordingly, identification and characterisation of these antibodies is relevant. The selected peptide could be subsequently used in earlier stage patients for the development of a novel and reliable diagnostic tool for anti-HNK-1 antibody identification in sera of patients affected by autoimmune neurological disorders monitoring disease activity
Fisher, Charles. "Statistical Characterization of Protein Ensembles". Thesis, Harvard University, 2012. http://dissertations.umi.com/gsas.harvard:10223.
Texto completoNiyogi, Sandip. "Synthesis and characterization of molecules to study the conformational barriers of fluorocarbon chains". Thesis, University of North Texas, 2000. https://digital.library.unt.edu/ark:/67531/metadc2511/.
Texto completoCherwa, Jr James Edward. "Characterization of Scaffolding Proteins Altered in the Ability to Perform a Critical Conformational Switch". Diss., The University of Arizona, 2009. http://hdl.handle.net/10150/195476.
Texto completoBraggin, Greg A. "Effect of Surfactant Architecture on Conformational Transitions of Conjugated Polyelectrolytes". DigitalCommons@CalPoly, 2015. https://digitalcommons.calpoly.edu/theses/1411.
Texto completoZhang, Xiaofan. "Synthesis, Characterization and Catalytic Studies of Chiral Gold Acyclic Diaminocarbene Complexes". Thesis, University of North Texas, 2016. https://digital.library.unt.edu/ark:/67531/metadc862827/.
Texto completoLee, Alexis J. "Theoretical Approaches to the Characterization of Water, Aqueous Interfaces, and Improved Sampling of Protein Conformational Changes". ScholarWorks@UNO, 2012. http://scholarworks.uno.edu/td/1511.
Texto completoPondaven, Simon Pierre. "Conformational Flexibility and Amyloid Core Characterization of Human Immunoglobulin Light Chain Domains by Multidimensional NMR Spectroscopy". The Ohio State University, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=osu1354113457.
Texto completoDollery, Stephen. "Identification and characterization of low pH-triggered conformational changes in the herpes simplex virus glycoprotein B". VCU Scholars Compass, 2011. http://scholarscompass.vcu.edu/etd/176.
Texto completoRaspadori, Andrea. "Characterization of the conformational space of the murine prion protein using single-molecule force spectroscopy techniques". Doctoral thesis, SISSA, 2014. http://hdl.handle.net/20.500.11767/4124.
Texto completoSymeonidou, Evgenia. "Synthesis, characterization and DFT study of new azaborinine compounds". Master's thesis, Alma Mater Studiorum - Università di Bologna, 2020. http://amslaurea.unibo.it/21700/.
Texto completoPosgai, Monica Therese. "Energetic and dynamic characterization of the IgA1:FcαRI interaction reveals long-range conformational changes in IgA1 upon receptor binding". University of Cincinnati / OhioLINK, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1354043317.
Texto completoZhao, Shen. "Design and in vitro characterization of lipids with a pH-sensitive conformational switch and their liposomes for anticancer drug delivery". Scholarly Commons, 2018. https://scholarlycommons.pacific.edu/uop_etds/3574.
Texto completoTESTA, LORENZO. "Conformational transitions of the intrinsically disordered protein sic1 from the yeast saccharomyces cerevisiae. Towards structural and functional characterization of the whibitory complex with CDK1-CLB5". Doctoral thesis, Università degli Studi di Milano-Bicocca, 2013. http://hdl.handle.net/10281/43673.
Texto completoMalloni, Wilhelm Massimiliano [Verfasser] y Hans Robert [Akademischer Betreuer] Kalbitzer. "AUREMOL-QTA, a program package for NMR based automated recognition and characterization of local and global conformational changes in proteins induced by ligand binding as external perturbation / Wilhelm Massimiliano Malloni. Betreuer: Hans Robert Kalbitzer". Regensburg : Universitätsbibliothek Regensburg, 2011. http://d-nb.info/1030178836/34.
Texto completoRauch, Sebastian, Klaus-Jochen Eichhorn, Ulrich Oertel, Manfred Stamm, Dirk Kuckling y Petra Uhlmann. "Temperature responsive polymer brushes with clicked rhodamine B: synthesis, characterization and swelling dynamics studied by spectroscopic ellipsometry". Saechsische Landesbibliothek- Staats- und Universitaetsbibliothek Dresden, 2014. http://nbn-resolving.de/urn:nbn:de:bsz:14-qucosa-139314.
Texto completoDieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich
Rauch, Sebastian, Klaus-Jochen Eichhorn, Ulrich Oertel, Manfred Stamm, Dirk Kuckling y Petra Uhlmann. "Temperature responsive polymer brushes with clicked rhodamine B: synthesis, characterization and swelling dynamics studied by spectroscopic ellipsometry". Royal Society of Chemistry, 2012. https://tud.qucosa.de/id/qucosa%3A27823.
Texto completoDieser Beitrag ist mit Zustimmung des Rechteinhabers aufgrund einer (DFG-geförderten) Allianz- bzw. Nationallizenz frei zugänglich.
Naidoo, Kevin Jonathan. "Metal ion cages with stabilized conformations : synthesis, characterization and properties". Master's thesis, University of Cape Town, 1989. http://hdl.handle.net/11427/18805.
Texto completoMossuto, Maria Francesca. "Protein amyloidogenesis: characterization of aggregation prone conformations and fibrils structure". Doctoral thesis, Università degli studi di Padova, 2008. http://hdl.handle.net/11577/3425566.
Texto completoKallberg, Yvonne. "Bioinformatic methods in protein characterization /". Stockholm, 2002. http://diss.kib.ki.se/2002/91-7349-370-8/.
Texto completoBanks, Jennifer Dawn. "Characterization of a minimal avian leukosis-sarcoma virus packaging signal /". Thesis, Connect to this title online; UW restricted, 1999. http://hdl.handle.net/1773/11528.
Texto completoPark, Sunho 1976. "Characterization of nanoparticle-DNA conjugate and control of DNA conformation on particle surface". Thesis, Massachusetts Institute of Technology, 2009. http://hdl.handle.net/1721.1/49761.
Texto completoIncludes bibliographical references.
Nano-science has exploited the hybridization and de-hybridization phenomena of DNA which are one of its fundamental functions. In particular, conjugates of gold nanoparticles and DNA (Au NP-DNA) have been extensively explored for their potential in biological applications such as DNA delivery for gene therapy and disease detection. However, DNA strands are known to adsorb onto the Au NP surface, which can severely limit the hybridization ability of Au NP-DNA conjugates. Therefore, methods of chemical modification of Au NP surfaces and evaluating DNA conformation via Ferguson analysis of gel electrophoresis are proposed in the thesis. Conjugates of DNA with Au NP of different sizes and coverages are evaluated with Ferguson analysis to characterize important parameters such as hydrodynamic size and zeta-potential. Surface modified Au NP exhibits enhanced stability and hybridization specificity in the system, which infers the effectiveness of those methods towards biological systems where non-specific adsorption is problematic. To confirm the validity of the concept, Au NP-antisense DNA experiments for gene silencing are performed in the work. Antisense DNA is designed to inhibit ribosomal activity on mRNAs and cooperatively works with Au NPs to enhance physical blocking mechanisms. However, the result shows that Au NP-DNA conjugates can enhance in vitro gene expression depending on DNA sequence and coverage of the conjugates. Suggestions are made for further investigation on proof and improvement of the translation enhancer concept.
by Sunho Park.
Ph.D.
Spuhler, Philipp S. "High-throughput detection of DNA orientation and conformation for characterization of protein-DNA interactions". Thesis, Boston University, 2012. https://hdl.handle.net/2144/31605.
Texto completoPLEASE NOTE: Boston University Libraries did not receive an Authorization To Manage form for this thesis or dissertation. It is therefore not openly accessible, though it may be available by request. If you are the author or principal advisor of this work and would like to request open access for it, please contact us at open-help@bu.edu. Thank you.
A deep understanding of disease processes requires understanding of DNA, RNA and protein function on a molecular level. Protein-DNA interactions play a crucial role in many of these processes, including in gene expression. The high-throughput capability of micro-array based technologies enables studies on the sequence dependence to determine the functional roles of protein-DNA binding. In vitro protein binding to double stranded DNA microarrays has proven to be an effective complementary technique to the in vivo methods, as this allows rapid characterization of the protein-DNA binding sequence specificity and much higher resolution of the binding site. In this dissertation, we apply Spectral Self-Interference Fluorescence Microscopy (SSFM) to develop a platform that improves upon existing in vitro protein binding microarray technologies. The platform permits measurement of the protein binding site without the need for protein tagging with radiolabels or fluorophores. The technology enables precise quantification of DNA conformation changes that are induced by protein binding. There are currently no viable high-throughput techniques available to investigate the sequence dependence on protein induced DNA bending. While they are not well understood, such structural changes are thought to play a critical role in transcription. A crucial component of the technique is the measurement of DNA orientation, as this permits the detection of the protein binding location and DNA conformation changes. We apply a novel polymeric scaffold to control the surface charge of the sensor and, thus, the orientation of surface immobilized oligonucleotides. The orientation is precisely quantified through sub-nanometer height localization of fluorophore labels on either end of the oligonucleotide probes. The DNA binding protein Integration Host Factor (IHF) is used as the model system to demonstrate a proof of principle for the platform. IHF is known to bind DNA with high sequence specificity and to induce a 160° bend. The binding position of IHF to the dsDNA probes is resolved to three nucleotides and the conformation changes that result due to a single nucleotide polymorphism in the consensus binding sequence are observed. The platform is scalable to permit observation of protein binding to hundreds of thousands ofunique DNA sequences on a single chip.
2031-01-01
Tseng, Hui-yuan [Verfasser] y Reinhard [Akademischer Betreuer] Fässler. "Identification and characterization of conformation-specific cytoplasmic integrin interactors / Hui-yuan Tseng ; Betreuer: Reinhard Fässler". München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2017. http://d-nb.info/1124395806/34.
Texto completoFarooqi, Mohammed Junaid. "METHODS FOR IN SITU PIEZOPHYSIOLOGICAL STUDIES: OPTICAL SECTIONING VIA STRUCTURED ILLUMINATION AND FLUORESCENCE BASED CHARACTERIZATION OF NADH CONFORMATION". Oxford, Ohio : Miami University, 2009. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=miami1249225952.
Texto completoXia, Nan. "Time-of-flight secondary ion mass spectrometry (ToF-SIMS) characterization of conformation and orientation of adsorbed protein films /". Thesis, Connect to this title online; UW restricted, 2003. http://hdl.handle.net/1773/9869.
Texto completoHolder, Isabelle T. [Verfasser]. "Non-Canonical Nucleic Acids in Bacteria -Structural Characterization and Functional Properties of Quadruplex and Triplex Conformations- / Isabelle T. Holder". Konstanz : Bibliothek der Universität Konstanz, 2015. http://d-nb.info/1110771606/34.
Texto completoMeyer, Gordon Joel. "Synthesis, Characterization, and Mixed-Valence Studies of Conformationally Constrained Bisferrocenyl Complexes for the Study of Through-Space S***π; Interactions". Diss., The University of Arizona, 2014. http://hdl.handle.net/10150/337289.
Texto completoAgrahari, Aditya. "Synthesis and Characterization of Di- Aryl Pentanes and Mechanistic Study of Aldol Reaction of 9-Acetylanthracene with Paraformaldehyde". Cleveland State University / OhioLINK, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=csu1440082002.
Texto completo[Verfasser], Felix Helge Deluweit y Christiane [Akademischer Betreuer] Ritter. "Conformation based in vitro selection of mammalian prion seeds – Propagation and characterization of prion strains / Felix Helge Deluweit ; Betreuer: Christiane Ritter". Braunschweig : Technische Universität Braunschweig, 2015. http://d-nb.info/1175819646/34.
Texto completoHan, Zhong. "Characterization of the mechanisms of transcription termination by the helicase Sen1". Thesis, Université Paris-Saclay (ComUE), 2017. http://www.theses.fr/2017SACLS202/document.
Texto completoPervasive transcription is a common phenomenon both in eukaryotes and prokaryotes that consists in the massive production of non-coding RNAs from non-annotated regions of the genome. Pervasive transcription poses a risk that needs to be controlled since it can interfere with normal transcription of canonical genes. In S.cerevisiae, the helicase Sen1 plays a key role in restricting pervasive transcription by eliciting early termination of non-coding transcription. Sen1 is highly conserved across species and mutations in the human Sen1 orthologue, senataxin (SETX), are associated with two neurological disorders. Despite the major biological relevance of Sen1 proteins, little is known about their biochemical properties and precise mechanisms of action. During my PhD I have studied in detail the mechanisms of termination by Sen1.In a first project, I have characterized the biochemical activities of Sen1 and investigated how these activities partake in termination. To this end I have employed a variety of in vitro approaches, including a minimal transcription-termination system containing only purified Sen1, RNA polymerase II (RNAPII) and DNA transcription templates that allows modifying the different elements of the system in a controlled manner to understand their role in termination. First, we have analysed the function of the different domains of Sen1 in termination. Sen1 is a large protein composed of a central catalytic domain flanked by additional domains with proposed roles in protein-protein interactions. We have demonstrated that the central helicase domain is sufficient to elicit transcription termination in vitro. Next, we have shown that Sen1 can translocate along single-stranded nucleic acids (both RNA and DNA) from 5’ to 3’. Then, we have analysed the role of the different nucleic acid components of the elongation complex (i.e. nascent RNA and DNA transcription templates) in termination. Our results indicate that termination does not involve the interaction of Sen1 with the DNA but requires Sen1 translocation on the nascent RNA towards the RNAPII. Importantly, we show that upon encountering RNAPII, Sen1 can apply a mechanical force on the polymerase that results in transcription termination when RNAPII is paused under certain conditions. This indicates that RNAPII pausing is a strict requirement for Sen1-mediated termination. In a second project, in collaboration with the group of E. Conti we have performed a structure-function analysis of the helicase domain of Sen1. Comparison of Sen1 structure with that of other related helicases has revealed an overall similar organization consisting in two tandem RecA-like domains from which additional accessory subdomains protrude. In general, the core RecA-like domains are very well conserved among related helicases and most variation is found in the accessory subdomains, that often confer specific characteristics to different helicases. Indeed, we have found that Sen1 contains a unique but evolutionary conserved structural feature that we have dubbed the “brace”. In addition, Sen1 is different from other helicases in an auxiliary subdomain that we have named the “prong”. Importantly, we have shown that the integrity of this subdomain is critical transcription termination by Sen1. We propose that the specific features identified in our structural analyses are important determinants of the transcription termination activity of Sen1. Finally, we have used Sen1 as a model to investigate the molecular effect of SETX mutations linked to neurodegenerative diseases. We have introduced disease-associated mutations in Sen1 and performed a complete biochemical characterization of the different mutants in vitro. Importantly, we found that all mutants were severely affected in transcription termination. Taken together, our results elucidate the key structural determinants of the function of Sen1 and shed light on the molecular origin of the diseases associated with SETX mutations
Raphela, Mashikoane Pinky Jane. "Molecular characterization of Mycoplasma synoviae in chickens in South Africa using single-stranded conformation polymorphism and high-resolution melting curve analysis of the vlhA gene". Diss., University of Pretoria, 2012. http://hdl.handle.net/2263/26196.
Texto completoDissertation (MSc)--University of Pretoria, 2012.
Veterinary Tropical Diseases
unrestricted
Francou, Bruno. "Contribution à la caractérisation de nouveaux gènes impliqués dans les hypogonadismes hypogonadotropes : caractérisation des mécanismes moléculaires et cellulaires". Thesis, Université Paris-Saclay (ComUE), 2016. http://www.theses.fr/2016SACLS101/document.
Texto completoCongenital hypogonadotropic hypogonadism (CHH) is characterized by deficient or absent pubertal development due to deficient or absent secretion of the pituitary gonadotropins. The many known genetic causes are generally classified into distinct nosological groups. One comprises abnormalities that affect the pre-natal development or migration of GnRH neurons, the paradigm of which is Kallmann syndrome. The other encompasses molecular abnormalities that only affect hypothalamic GnRH synthesis, GnRH release or GnRH signaling at pituitary level. At this stage, two populations of hypothalamic neurons implicated in a gonadotrop function are identified, GnRH neurons and KNDy neurons secreting kisspeptins and neurokinin B. All of the identified genes would represent less than 20% of genetic etiologies.The aim of this PhD was to study the prevalence and pathophysiology mechanisms of known genes and to identify new genetic etiologies of CHH.In the first part, we characterized the function of all molecular events identified on KISS1R, TACR3 and TAC3 genes. Prevalences were estimated in 600 patients. A particular neuroendocrine profile was identified in patients presenting an alteration of neurokinin B signaling. Importance of Kisspeptins during embryonic life was validated. According to these data, a model of interaction between GnRH and KNDy neurons was proposed.In the second part, we identified two new CHH genes using various molecular genetics approaches. SEMA3A was identified in a familial form of Kallmann syndrome and PNPLA6 in a rare familial form of CHH.Finally, our increased knowledge of the various genetic forms of CHH allows proposing a new genetic approach based on next generation sequencing to test together all known and several candidate genes
Coty, Jean-Baptiste. "Caractérisation des nanomédecines pour la clinique : développement de méthodes évaluant les interactions nanoparticules-protéines plasmatiques pour une application en contrôle qualité". Thesis, Université Paris-Saclay (ComUE), 2017. http://www.theses.fr/2017SACLS432/document.
Texto completoNanomedicines injected intravenously interact with surrounding biological elements in the bloodstream. Among these interactions, those with blood proteins turn out to be very important regarding the becoming of the nanovectors. They acquire a biological identity upon interaction with proteins which influence their path to target tissue and cells. The understanding and mastering of these phenomena remains a crucial issue in nanomedicine development. Methods allowing an easier study of these interactions are needed. The aim of these PhD thesis was to develop such methods, usable on a routine basis in a clinical context, allowing a fine characterization of nanomedicines and their interactions with plasmatic proteins. This PhD is part of the project “Nano Innovation for CancEr” (NICE, BPI France), gathering a consortium of industrials partners developing clinical nanomedicines.In a first time, a bibliographic study about current methods used for such a characterization could identify two major limitations. (i) On one hand, the complexity of current available methods for which the equipment specificity and required expertise prevent their use at a large scale. (ii) On the other hand, properties today characterized on a daily basis (size, morphology, charge) are too rough compared to the sharpness of biological processes who interact and “analyze” the nanovectors introduced in biological media. These two aspects are limiting a safer development of nanomedicines as well as a good reproducibility of their action in clinics.During this thesis, we developed methods allowing a beginning of answer to the wide problematic of nanomedicine characterization. A method for a high throughput analysis of complement activation by nanomedicines via 2D immunoelectrophoresis was developed and validated. It allows the reproducible analysis of protein C3 fragmentation. This method is applicable to the study of the impact of nanoparticles in human serum and their degree of action on the complement cascade. This method has been used for a more fundamental study on complement activation pathways activated according to the architecture of nanoparticles surface.A second method for the study of complement activation produced by nanoparticles has been proposed using surface plasmon resonance (SPR). A chip allowing an automated screening of complement activation has been developed. This method was compared to other methods for complement activation study (2D immunoelectrophoresis, ELISA) and allowed the identification of bias during nanomedicine evaluation.Finally, an original approach for the characterization of nanomedicine’s surface architecture using proteins as molecular probes has been proposed. In this method, capillary electrophoresis has been used as analytical tool to allow a direct analysis of sample without preliminary nanoparticle removal step.Methods developed during this work can be applied to the characterization of nanomedicines and proposed as routine methods for quality control. A development of nanomedicines characterization in this direction constitute one of the lever for a more fruitful translation of nanomedicines entering in clinical phase
Fleischmann, Matthias [Verfasser], Ruth M. [Akademischer Betreuer] Gschwind, Kirsten [Akademischer Betreuer] Zeitler y Manfred [Akademischer Betreuer] Scheer. "NMR investigations on catalysts and conformations in organo- and photocatalytic reactions, and characterization of electrolytes and supramolecular switchable container molecules / Matthias Fleischmann. Betreuer: Ruth M. Gschwind ; Kirsten Zeitler ; Manfred Scheer". Regensburg : Universitätsbibliothek Regensburg, 2011. http://d-nb.info/1026165520/34.
Texto completoXiao, Jianxi. "NMR conformational and dynamic characterization of triple helical peptides". 2009. http://hdl.rutgers.edu/1782.2/rucore10001600001.ETD.000051923.
Texto completoWeng, Shao-En y 翁紹恩. "Conformational and dynamic characterization of Na+-dependent bile acid transporter ASBTNm". Thesis, 2018. http://ndltd.ncl.edu.tw/handle/q2dsnz.
Texto completo國立中興大學
生物化學研究所
106
Apical Sodium-dependent Bile acid Transporter (ASBT), a Na+-dependent secondary active transporter, is localized at the apical membrane of enterocytes and mediates the reuptake of bile acid from ileum back to liver. The structural information of ASBT is attributed to the crystal structures of bacterial homologs ASBTNm and ASBTYf, revealing the snapshots at inward- and outward-facing states, and the putative mechanism of substrate translocation via an “elevator-like” movement. Nevertheless, the triggers of conformational changes and the dynamics at different substrate conditions remain undetermined. In this project we employed Site-directed Alkylation monitored by in gel Fluorescence (SDAF) to probe the in situ alteration of substrate binding pocket accessibility. We tried to apply SDAF assay in topology determination of membrane proteins, and the results demonstrated it is valid and much more efficient than previous methods. In our earlier SDAF studies, a series of residues located at the substrate pathway in ASBTNm were substituted with cysteine, but the functional effects of these point mutations are still unknown. We performed whole cell [3H]-taurocholate uptake assay using ASBTNm-overexpressed E. coli C43 to monitor the transportation activities of these mutants. Although most ASBTNm mutants have lower activities than WT ASBTNm, they still reserve transport activity at certain level. In order to validate our dynamics data obtained by SDAF, we spin-labelled Y39 and T303 for Double Electron-Electron Resonance (DEER) using Electroparamagnetic Resonance (EPR) spectroscopy. The DEER results indicated that DDM-solubilized ASBTNm is populated as two discrete conformational species in K+ buffer, representing outward- and inward-facing states, and one major conformational species in Na+ buffer, and in taurocholate added Na+ buffer, representing inward-facing and occluded states. In order to obtain soluble ASBTNm in lipid bilayer for DEER spectroscopy, we tried to purify ASBTNm in styrene maleic acid lipid particles (SMALP). The C-terminal GFP fusion facilitates tracking of ASBTNm-SMALP, although the purity requires further optimization. The soluble membrane protein particles in SMALP will enable us to preserve the native behavior of ASBTNm for structural and dynamical studies. We also analized surface charge of ASBTNm using APBS, and performed SwissDock for prediction of possible substrate binding sites. The predicted results are of our expectation. The studies will provide practical methods in obtaining crystal structure of ASBTNm bound with taurocholate at outward-facing state.
Simão, Alcides Pinto. "Gas-Phase Conformational Characterization of Trace Amine Neurotransmitters: a rotational spectroscopy study". Doctoral thesis, 2020. http://hdl.handle.net/10316/91053.
Texto completoA presente dissertação trata das preferências conformacionais de uma série de aminas residuais (octopamina, sinefrina, fenilglicinol e fenilefrina), bem como de um precursor aminoácido (ácido pipecólico) e de um aminossacarídeo (glucosamina), estudadas em condições de isolamento em fase gasosa recorrendo ao uso de uma combinação de técnicas de espectroscopia rotacional hifenada com ablação laser. Livre de qualquer tipo de interacção intermolecular, torna-se possível a caracterização f física das interacções intramoleculares para cada confórmero, bem como o seu papel no panorama conformacional de cada biomolécula. Os resultados experimentais foram posteriormente comparados com os resultados teóricos, de modo a se proceder a uma caracterização completa dos confórmeros observados. Esta correlação de dados permitiu ainda estabelecer a natureza das principais interacções intramoleculares envolvidas em cada conformação.
In the present dissertation, the conformational preferences of a series of trace amines (octopamine, synephrine, phenylglycinol and phenylephrine), as well as an aminoacid precursor (pipecolic acid) were studied in gas phase isolation conditions using a combination of rotational spectroscopy tech- niques together with laser ablation. Free from any kind of intermolecular interaction, the physical characterization of the intramolecular interactions for each conformer and their role in the conformational panorama of each biomolecule became possible. Experimental results were then compared with theoretical results, in order to fully characterize the observed conform- ers. This data correlation also allowed to establish the nature of the major intramolecular interactions involved in each conformation.
Hsiao, Chun-Che y 蕭均哲. "Characterization of the conformational and functional properties of two in vitro refolded Bacillus subtilis SigB". Thesis, 2016. http://ndltd.ncl.edu.tw/handle/73692598406431862636.
Texto completo國立中興大學
生物化學研究所
104
Transcription initiation is the first step of gene expression and a major point for gene regulation.σ factor of RNA polymerase (RNAP;α2ββ''σ) confers the specificity of promoter recognition and initiates transcription. It is also involved in open complex, abortive transcription and regulation of gene expression. Recent works in our lab demonstrated that the N-terminally truncated primary σA factor, SND100-σA, and σB of Bacillus subtilis, both of which lack region 1.1, are able to specifically interact with promoter DNA in the absence of core RNA polymerase in vitro. Moreover, SND100-σA factors with different functional properties were achieved through the use of slightly different refolding protocols. Among them, SND100-σA1 is capable of exhibiting specific and core RNAP-independent promoter-binding activity; whereas SND100-σA3 is able to melt DNA independent of core RNA polymerase in vitro. Present study is aimed to investigate whether the B. subtilis σB can also adopt different functional properties through protein refolding. To fulfill this goal, the σB protein was overexpressed in E. coli BL21 (DE3) harboring a σB-expressing plasmid. The overexpressed σB in inclusion bodies were denatured with guanidine hydrochloride and refolded through mild serial dilution of the guanidine hydrochloride solution. The refolded σB were soluble and can be purified using Superdex G-200 and Superdex G-75 columns. Buffers with or without both PMSF and DTT were used for protein refolding and purification and the σB proteins thus obtained were named σB1 and σB3, respectively. Results of circular dichroism reveal that σB1 and σB3 have similar contents of secondary structures albeit with different overall conformations. Moreover, both σB1 and σB3 are able to bind promoter DNA as analyzed by electrophoretic mobility shift assay (EMSA). However, the two σB-promoter DNA complexes are different in sensitivity to heparin challenge, suggesting that they possesss different DNA-binding properties.
Baldwin, Richard Kimo. "Preparation, characterization and conformational variability of [eta]6-Hexaethylbenzene and other arene complexes of ruthenium". Phd thesis, 1998. http://hdl.handle.net/1885/144675.
Texto completoZHANG, JIANMING. "Conformational and Mechanical Characterization of Organic Thin Films on Surfaces by Neutron Reflection and Atomic Force Microscopy". Diss., 2014. http://hdl.handle.net/10161/9400.
Texto completoEngineering thin, organic materials with tailored properties requires both the understanding of the conformation of thin organic films and their conformational response to changes in the environment, and the accurate characterization the mechanical properties of the materials as a thin layer on surfaces. These issues have not yet been sufficiently addressed due to the paucity of appropriate tools and data interpretation approaches to reveal the nanometer scale conformation and mechanics of surface-grafted, thin, organic films. In this dissertation, I report on the characterization of conformational and mechanical properties of thin organic films, and the development of techniques that allow more detailed and reliable measurement of these material properties. First, I co-developed a novel approach to evaluate neutron reflectivity data and to simulate the conformational structure for thin stimulus-responsive polymer brushes. In this approach, we used a molecular-based lattice mean-field theory, augmented with experimentally obtained parameters to describe the polymer chains. The approach and fitting results required fewer fitting parameters, and captured the thermal response of the sample self-consistently.
Second, I demonstrated the capability of force-modulation microscopy in imaging surface-grafted, organic thin films in aqueous environments, with high spatial resolution and sensitivity to conformational details that affect the contact mechanics. To this end, I developed a new parameter-selection approach. This approach allowed both highly sensitive mapping of subtle differences in the molecular packing of thiol molecules on the substrate surface, and generation of high-contrast contact-stiffness images of end-grafted protein patterns on a surface. Finally, I discussed model selection and error estimation in calculating the reduced Young's modulus of soft materials on surfaces. I found that the detailed characterization of probe apex profiles, using probe-reconstruction techniques, provide only marginal improvements in calculating the reduced Young's modulus of thin films, compared with analytical models of equivalent probe radii; however, I found that a hybrid worn-cone model is appropriate for large indentations on soft materials, and benefits from the characterization of the probe apex profile. Additionally, we rendered error maps of several common scenarios, referenced to indentation and probe radius values, in the determination of the reduced Young's modulus.
Dissertation
Hansen, Jeffrey Craig. "Characterization of the conformational transitions of the estrogen receptor monomer by partition in aqueous two-phase systems". 1986. http://catalog.hathitrust.org/api/volumes/oclc/15244978.html.
Texto completoTypescript. Vita. eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references (leaves 153-162).
Ujvari, Andrea. "Characterization of DNA structural elements and conformational changes accompanying the binding of T7 RNA polymerase to its promoter". 1999. https://scholarworks.umass.edu/dissertations/AAI9920662.
Texto completoTeklebrhan, Robel Berhe. "Conformational preferences and structural characterization of prolyl cis/trans isomerization of carbohydrate-templated proline mimetics of some model peptides using computational methods". 2009. http://hdl.handle.net/1993/21590.
Texto completoTseng, Rong-Jeng y 曾榮正. "(1) Study of The Regulation of the CFTR Channel Gating(2) Characterization of Mg2+, K+ and ATP Induced Conformational Change of Yeast Hsp60". Thesis, 1995. http://ndltd.ncl.edu.tw/handle/73778083115452467256.
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