Дисертації з теми "Deoxy"
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Roig, Ricard. "Synthesis of 6-deoxy-6-fluorosugars." Thesis, University of Leicester, 2006. http://hdl.handle.net/2381/29987.
Повний текст джерелаPark, Sung-Hae. "The biosynthesis of the deoxyhexose moieties in oleandomycin /." Thesis, Connect to this title online; UW restricted, 1999. http://hdl.handle.net/1773/8152.
Повний текст джерелаSande, Marc van de. "Asymmetrische Synthese von 3-Oxa-15-Deoxy-16-(m-tolyl)-Tetranorisocarbacyclin und 15-Deoxy-16-(m-tolyl)-Tetranorisocarbacyclin." Aachen Mainz, 2007. http://d-nb.info/100174103X/34.
Повний текст джерелаFazio, Fabio. "Building blocks for 2-deoxy-L-nucleosides." [S.l. : s.n.], 2001. http://deposit.ddb.de/cgi-bin/dokserv?idn=963273434.
Повний текст джерелаJiao, Hailong. "Synthetic studies on 2-amino-2-deoxy glycosides." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1999. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape7/PQDD_0002/NQ39546.pdf.
Повний текст джерелаEllenberger, Suzanne Ray. "Total synthesis of selected deoxyamino sugars /." Full text open access at:, 1986. http://content.ohsu.edu/u?/etd,110.
Повний текст джерелаSande, Marc van de [Verfasser]. "Asymmetrische Synthese von 3-Oxa-15-Deoxy-16-(m-tolyl)-Tetranorisocarbacyclin und 15-Deoxy-16-(m-tolyl)-Tetranorisocarbacyclin / vorgelegt von Marc van de Sande." Aachen : Mainz, 2007. http://d-nb.info/100174103X/34.
Повний текст джерелаHeß, David. "Palladium(II) complexes and phenylboronic acid esters of deoxy sugars." Diss., lmu, 2012. http://nbn-resolving.de/urn:nbn:de:bvb:19-151675.
Повний текст джерелаHardy, Simon. "Approaches towards the total synthesis of the 20-deoxy bryostatins." Thesis, University of Manchester, 2009. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.496225.
Повний текст джерелаSteer, Andrew Mark. "Studies on the prebiotic origin of 2-deoxy-D-ribose." Thesis, University of York, 2017. http://etheses.whiterose.ac.uk/19070/.
Повний текст джерелаRose, Graeme William. "The synthesis of some 3-deoxy-α-keto sugar acids". Thesis, University of Edinburgh, 1988. http://hdl.handle.net/1842/14324.
Повний текст джерелаZhu, Danyang. "Stereoselective Synthesis of 2-Deoxy Glycosides via Anomeric O-Alkylation." University of Toledo / OhioLINK, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=toledo1438900175.
Повний текст джерелаCumming, Hemi Adam. "Probing the Substrate Specificity of 3-Deoxy-D-arabino-Heptulosonate 7-Phosphate Synthase and 3-Deoxy-D-manno-Octulosonate 8-Phosphate Synthase using Analogues of Phosphoenolpyruvate." Thesis, University of Canterbury. Chemistry, 2007. http://hdl.handle.net/10092/2190.
Повний текст джерелаKoo, Ki Chul. "Studies towards the total synthesis of 1-deoxy-8-demethyl taxol." Tallahassee, Florida : Florida State University, 2009. http://etd.lib.fsu.edu/theses/available/etd-08222009-032151/.
Повний текст джерелаAdvisor: Robert A. Holton, Florida State University, College of Arts and Sciences, Dept. of Chemistry and Biochemistry. Title and description from dissertation home page (viewed on April 27, 2010). Document formatted into pages; contains xvii, 362 pages. Includes bibliographical references.
Rodríguez, Gómez Miguel Angel. "Stereoselective Synthesis of 2-Deoxy-glycosides. Approach to the Synthesis of Digitoxine." Doctoral thesis, Universitat Rovira i Virgili, 2007. http://hdl.handle.net/10803/9010.
Повний текст джерела2-Deoxy and 2,6-dideoxy-glycosides are important structural units in many natural products including antitumor drugs (anthracyclines, aureolic acids, calicheamicin, esperamicin), antibiotics active against Gram-positive bacteria (erythromycins, orthosomycins), antibiotics inhibiting platelet aggregation (angucyclines), drugs used in the treatment of cardiac insufficiency (cardiac glycosides), antiparasitic agents (avermectins).
The stereocontrolled formation of the glycosidic linkage in 2-deoxy-oligosaccharides has been found to be one of the most challenging tasks in glycosylation reactions. This could be solved using C-2 substituents (I, SeR, SR) in order to aid stereocontrol in the glycosylation step. These groups can be then easily removed under mild conditions after stereocontrol has been achieved. On the other hand, one of the most important intermediates in the synthesis of 2 deoxycarbohydrates are 1,2-unsaturated glycosides, commonly called glycals.
The aim of the study reported in this thesis was to develop a new method for the stereoselective synthesis of 2-deoxyoligosaccharides and precursors such as glycals. Moreover, an approach to the synthesis of digitoxine was also reported. Thus, in a preliminary introduction, previous reported methods for the synthesis of deoxysugars were discussed. In chapter 1, we developed a new procedure for synthesizing phenyl 2-deoxy-2-iodo-1-thio-glycosides and their use as glycosyl donors for the stereocontrolled synthesis of 2-deoxy-2-iodo-oligosaccarides. The key step for the synthesis of 2-deoxy-2-iodo-1-thio-glycosides is a regio- and stereoselective 6-endo cyclization of alkenols induced by iodine electrophiles. In chapter 2, Deoxy-2-iodopyranosides were synthesized from sulfanyl alkenes using a "one pot" consecutive cyclization-glycosylation process. The "one pot" procedure was also applied to the synthesis of a 2,6-dideoxy-2-iodo-glycoside, which was successfully deiodinated to afford the 2,6-dideoxyglycoside. In chapter 3, it is showed that pyranoid glycals of all configurations can be obtained from pentoses through an olefination-cyclization-elimination sequence. This method provides access to non conventional glycals having C3-alkoxy substituent in axial configuration, such as D-allal and D gullal, which are difficult to synthesize by usual methods. Furthermore, other functionalized glycals such as 2 phenylselenenyl or 2 iodoglycals can be synthesized starting from enolthioethers by direct selenium-mediated elimination or through dehydrative reaction of 2-iodolactols, respectively. Finally, Starting from a common precursor such as D-ribonolactone, we have explored a new approach to the synthesis of digitoxine and other cardiac glycosides as application of the previous methodology developed.
Títol: "Stereoselective Synthesis of 2-Deoxyglycosides. Approach to the synthesis of Digitoxine"
Paraules Clau: Síntesi estereoselectiva, 2-desoxi i 2,6-didesoxiglicòsids, glicósids cardiacs, glicals i iodoglicals, síntesi "one-pot", ciclació (electròfila), digitoxina.
Informe Preceptiu sobre la Tesi doctoral "Estereoselective Synthesis of 2 Deoxyglycosides. Approach to the síntesis of Digitoxine" presentada per Miguel Àngel Rodríguez Gómez.
La tesi s'emmarca dins el camp de la síntesis de carbohidrats i glicoconjugats amb estructures que presenten posicions on una o més de les funcions hidroxil característiques del sacàrids no hi estan presents. El treball realitzat ha tingut com objectiu final la síntesis estereoselectiva de 2-desoxi i 2,6-didesoxiglicòsids ja que són part constituent de moltes substàncies biològicament actives i/o productes naturals com antitumorals, antibiòtics, agents antiparasitaris, cardiotònics...i a més a més són difícils d'obtenir a partir de carbohidrats naturals.
D'aquesta forma en aquesta tesi s'aborda la síntesis de 2-desoxi-2-iodo-1-tiopiranósids com a nous dadors de glicosil i la seva aplicació en la síntesis estereoselectiva d'oligosacàrids i glicòsids. Aquest dadors de glicosil es caracteritzen per la presència d'un grup fenilsulfanil com a grup sortint en la posició anomèrica (C1) i un grup iodo en el C2 que actua com element de control en la reacció de glicosilació. Aquests dos grups funcionals donen, a més a més, moltes possibilitats de derivatització i una variada reactivitat.
La memòria s'ha organitzat en una introducció general sobre la biologia i la química dels 2-desoxi i 2,6-didesoxiglicòsids, un objectius, quatre capítols on es s'exposen i discuteixen els resultats obtinguts amb les seves corresponents conclusions i un annex amb els espectres dels productes seleccionats.
La introducció tracta sobre la importància i el variat paper biològic dels 2-desoxi i 2,6-didesoxicarbohidrats a la vegada que parla de la dificultat i els especials problemes que comporta la síntesis química d'aquests tipus de compostos. D'aquesta forma es fa una revisió dels mètodes desenvolupats fins avui per la síntesis d'aquest glicòsids. Lligat amb aquests mètodes anteriors, en els objectius es posa de manifest la necessitat d'arribar a un nou mètode de síntesis de 2-desoxicarbohidrats que permeti assolir totes les configuracions de piranòsids possibles.
En el primer capítol es desenvolupa el nou mètode d'obtenció de 2-desoxi-2 iodo-1-tioglicòsids que després es faran servir com a dadors de glicosil. D'aquesta forma, partint de pentoses de totes les configuracions i diferentment protegides es van olefinar per diversos mètodes, obtenint polihidroxihexenilsulfurs. El mètode més convenient per aquesta reacció en termes de rendiment i estereoselectivitat fou la olefinació amb oxid de fosfina (Wittig-Horner-WH). Aquests alquenols es van ciclar amb electrofils de iode, conduint de forma regioselectiva als 2-desoxi-2-iodo-1 tiopiranòsids. Aquest dadors de glicosil es van fer reaccionar amb colesterol com a model d'aglicona de diferents compostos bioactius i amb un glucosid com a model de síntesi d'oligosacàrid.
En el segon capítol s'aborda la síntesis dels mateixos compostos del capítol primer aprofitant la semblança de les condicions de ciclació i de glicosilació, que permet en una sola etapa la glicosilació de diversos compostos partint de l'alquenol, un precursor acíclic molt més estable i fàcil de sintetizar que el 2-desoxi-2-iodo-tioglicòsid corresponent. Aquest procediment "one-pot" es mostra igual en diasteroselectivitat i superior en termes de rendiment i de facilitat de manipulació que la síntesi per passos del capítol 1. A més es va sintetizar un 2,6-didesoxicarbohidrat model del supressor de l'apetit P57AS3.
En el tercer capítol s'exposa la síntesi de glicals a partir del 1-tio-2-desoxi-2 iodo-piranosids. El glicals són compostos molt versàtils i útils en la síntesis de carbohidrats i amb el procediment desenvolupat en aquest capítol s'arribà a obtenir glicals de configuracions difícils d'obtenir per altres mètodes, com el D-allal i el D gullal. A més, en una segona part del capítol tercer, aplicant un procediment de glicosilació estàndar per a com el de Gin ("dehydrative glycosylation") s'obtenen a partir de 2 iodolactols diversos compostos com a 2 iodoglicals, glicals o 1,1'-disacàrids.
En el quart capítol tots els anteriors procediments s'apliquen en la aproximació a la síntesis d'un glicósid cardíac, la digitoxina, molt utilitzat en el tractament de la insuficiència cardíaca. D'aquesta forma es realitzà la síntesi dels alquenols precursors dels 2,6-dideoxiglicòsids que formen part de l'estructura d'aquest fàrmac (unitats de digitoxosa) i es va unir a la aglicona obtenint el monosacàrid de la digitoxigenina. A més a més es van obtenir altres intermedis valuosos, tals com el corresponents 2 iodolactols o els trichloroacetimidats, en el camí cap a la síntesis de la digitoxina i/o anàlegs.
Amb el treball d'aquesta tesi els objectius inicialment proposats de desenvolupament d'un nou mètode de glicosilació per la síntesi de 2-desoxiglicòsids i han estat ampliament assolits i a més a més, s'han desenvolupat vies alternatives (glicals, lactols, 2-iodoglicals,...) que amplien els procediments sintètics inicials, ampliant les vies de síntesis dels 2-desoxiglicòsids.
Schell, Christoph. "Steuerung humaner testikulärer peritubulärer Zellen durch TNFalpha und 15-deoxy-Prostaglandin J2." Diss., lmu, 2012. http://nbn-resolving.de/urn:nbn:de:bvb:19-142254.
Повний текст джерелаEbel, Susanne. "The synthesis and study of oligo(deoxy)ribo-nucleotides and their analogues." Thesis, University of Edinburgh, 1993. http://hdl.handle.net/1842/13766.
Повний текст джерелаPongdee, Rongson. "New methods for 2-Deoxy-Beta-Oligosaccharide synthesis and progress towards the total synthesis of Lomaiviticinone." Texas A&M University, 2003. http://hdl.handle.net/1969.1/356.
Повний текст джерелаMcCarter, John D. "Deoxy and deoxyfluoro glycosides as mechanistic probes of Escherichia coli (lacZ) B-galactosidase." Thesis, University of British Columbia, 1991. http://hdl.handle.net/2429/30083.
Повний текст джерелаScience, Faculty of
Chemistry, Department of
Graduate
Harrison, Aidan Nicholas. "Investigations into the Inhibition of 3-Deoxy-D-manno-Octulosonate 8-Phosphate Synthase." Thesis, University of Canterbury. Chemistry, 2010. http://hdl.handle.net/10092/5382.
Повний текст джерелаLiang, Theresa. "Silver-Mediated Trifluoromethoxylation of Aryl Nucleophiles and Synthesis of 3-Deoxy-3-Fluoromorphine." Thesis, Harvard University, 2012. http://dissertations.umi.com/gsas.harvard:10547.
Повний текст джерелаChemistry and Chemical Biology
Englert, Nadine Esther [Verfasser]. "NMR Strukturanalyse der 1-Deoxy-D-xylulose-5-phosphat Reduktoisomerase / Nadine Esther Englert." Gießen : Universitätsbibliothek, 2012. http://d-nb.info/1064837557/34.
Повний текст джерелаWhite, Justin K. "Investigations into the Non-Mevalonate Isoprenoid Biosynthesis Pathway's First Two Enzymes utilizing Hybrid QM/MM Techniques." Scholar Commons, 2017. http://scholarcommons.usf.edu/etd/7107.
Повний текст джерелаAstles, David James. "Synthetic and mechanistic studies in the field of deoxy, branched-chain and amino sugars." Thesis, University of Ottawa (Canada), 1985. http://hdl.handle.net/10393/4768.
Повний текст джерелаZhu, Jiang. "Synthesis and mechanistic studies on 2-deoxy-Ã- and ß-D-glucopyranosyl pyridinium tetrafluoroborates." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 2000. http://www.collectionscanada.ca/obj/s4/f2/dsk1/tape3/PQDD_0027/NQ51942.pdf.
Повний текст джерелаBierau, Jörgen. "The pivotal role of CTP synthetase in the metabolism of (deoxy)nucleosides in neuroblastoma." [S.l. : Amsterdam : s.n.] ; Universiteit van Amsterdam [Host], 2003. http://dare.uva.nl/document/70985.
Повний текст джерелаIsaac, Siara. "Synthesis and gene silencing activity of ribonucleic acid duplexes containing 3'-deoxy-3'-thiothymidine." Thesis, McGill University, 2009. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=32408.
Повний текст джерелаCette thèse présente la première synthèse et caractérisation biophysique d'oligoribonucleotides (ARN) contenant des unités de 3'-deoxy-3'-thiothymidine (3'-S-dT). Des conditions optimisées pour l'intégration de 3'-S-dT en 21-nt ARN sont aussi présentées. L'impact de la substitution de 3'-S-dT sur la structure et la stabilité des duplex ARN:ARN fut recherché systématiquement par l'entremise de dichroïsme circulaire (CD) et d'études UV/thermale (Tm). Généralement, le remplacement du 3'-oxygène par un atome de sulfure résulta en une baisse significative de la stabilité thermique du duplex, et des effets de positionnement important furent observés. Aucune disruption significative de la structure hélicoïdale A-form du duplex ne fut détectée par le CD, quoi que parfois des inserts de 3'-S-dT entraînèrent une réduction des interactions base-base et/ou une distorsion locale du duplex. Des essaies révélèrent que des duplex interférents d'ARN court (siRNA) contenant du 3'-S-dT sur le brin guide n'ont pas baissé la production de gènes, mais que celles contenant du 3'-S-dT sur le brin complémentaire OU sur les deux brins présentèrent des activités qui fréquemment surpassaient celles de duplex non-modifiés de siRNA. Les substitutions à la position phosphate de rupture du brin complémentaire suggèrent que l'étape déterminante du clivage par RISC n'implique pas le 3'-oxygène pour la coordination d'un ion de métal.
Wilds, Christopher James. "Synthesis, physicochemical and biological properties of oligonucleotides containing 2-fluoro-2-deoxy-℗-D-arabinose." Thesis, McGill University, 1999. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=36730.
Повний текст джерелаAll four nucleobase monomers (thymine, cytosine, adenine and guanine) were prepared for solid phase oligonucleotide synthesis, from which homopolymeric and heteropolymeric base sequences were assembled. These molecules bound with very good affinities to both DNA and RNA targets. Structural studies via NMR experiments demonstrated that in a 2'F-ANA/RNA duplex the 2'F-ANA residues adopt an O4' -endo conformation, similar to what has been proposed for the structure of DNA in a DNA/RNA hybrid.
A 2'F-ANA oligopyrimidylate formed a triple-helical complex with duplex DNA and hybrid DNA(Pu):RNA(Py) with an affinity higher than that of a corresponding DNA strand. Also, a cytosine-rich 2' F-ANA strand was found to form a complex at acidic pH which has properties similar to that of i-motif DNA.
Finally, 2'F-ANA strands when hybridized to RNA were found to activate RNase H, an enzyme that is involved in the mechanism of action of antisense drugs. 2'F-ANA is the first example of an antisense analogue that demonstrates improved binding to RNA relative to DNA and still retains the ability to elicit RNase H activity.
Tran, David. "Investigating the substrate specificity of 3-deoxy-D-arabino-heptulosonate 7-phosphate (DAH7P) synthase." Thesis, University of Canterbury. Chemistry, 2011. http://hdl.handle.net/10092/6565.
Повний текст джерелаBlackmore, Nicola Jean. "The regulation of 3-deoxy-D-arabino-heptulosonate 7 phosphate synthase from Mycobacterium tuberculosis." Thesis, University of Canterbury. Chemistry, 2015. http://hdl.handle.net/10092/10242.
Повний текст джерелаReichau, Sebastian. "Inhibition and regulation of Mycobacterium tuberculosis 3-deoxy-D-arabino-heptulosonate 7-phosphate synthase." Thesis, University of Canterbury. Chemistry, 2013. http://hdl.handle.net/10092/7896.
Повний текст джерелаHellmuth, Isabell [Verfasser]. "Functionally modified (Deoxy)Ribonucleotides : synthesis and study of physicochemical and biological properties / Isabell Hellmuth." Mainz : Universitätsbibliothek Mainz, 2017. http://d-nb.info/1132364590/34.
Повний текст джерелаMehta, Dipti J. "15-deoxy-delta-12, 14-prostaglandin J2 (15d-PGJ2) Mediated Signaling in Colon Cancer." Diss., The University of Arizona, 2006. http://hdl.handle.net/10150/194039.
Повний текст джерелаZagorodna, Oksana. "Bcl-2 family members regulate the sensitivity to 2-deoxy-D-glucose in lymphomas." Diss., University of Iowa, 2011. https://ir.uiowa.edu/etd/2794.
Повний текст джерелаMontel, Sonia. "La 1-Deoxy-D-Xylulose-5-Phosphate Reductoisomerase, une métalloenzyme cible pour l'élaboration d'inhibiteurs chélatants." Thesis, Montpellier, Ecole nationale supérieure de chimie, 2012. http://www.theses.fr/2012ENCM0013.
Повний текст джерелаThe non-mevalonate pathway is highly present in higher plants, protozoa and bacteria but as no equivalent in mammals. That is why shut down isoprenoid biosynthesis and identify a non-mevalonate pathway inhibitor would greatly contribute to the search for safer antibiotics, antimalarials and for our concern herbicides. The unique properties of the 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR), the central enzyme of this pathway, make it a remarkable and attractive target for drug design. Fosmidomycin acts as an inhibitor of DXR and still remains, along with its N-acetyl homologue FR90098, one of the most potent inhibitor ever known even if extensive work on the development of Fosmidomycin analogue derivatives have been developed since the last decade as demonstrated in the first chapter with the development of a structure activity relationship of all the potential inhibitors of this enzyme already reported in the literature. The X-ray diffraction analysis of the co-crystals of DXR and Fosmidomycin or substrate shows that the phosphonic/phosphate group interacts with a highly specific polar pocket in the enzyme site, allowing only few structural modifications. By contrast, the cation chelating subunit represented by the hydroxamic acid function offers fine tuning possibilities for the complexation abilities as well as potential secondary interactions with the NADPH cofactor or directly with the enzyme. In this context, several modifications such as the introduction of carbamoylphosphinate, amidoxime, N-hydroxyurea and uracil complexing subunits have been made in order to find new families of DXR inhibitors. All of these functions show promising chelation capabilities as they already led to potent inhibitors of different metalloenzymes
Walker, Scott Raymond. "Design, Synthesis and Characterisation of Inhibitors of 3-Deoxy-D-arabino-Heptulosonate 7-Phosphate Synthase." Thesis, University of Canterbury. Chemistry, 2007. http://hdl.handle.net/10092/3932.
Повний текст джерелаCross, Penelope Jane. "Unravelling the Evolution of Allosteric Regulation in 3-Deoxy-D-arabino-heptulosonate 7-phosphate Synthase." Thesis, University of Canterbury. Chemistry, 2012. http://hdl.handle.net/10092/6823.
Повний текст джерелаOthman, Mohamad. "Characterisation and Control of 3-Deoxy-D-arabino-heptulosonate 7-phosphate Synthase from Geobacillus sp." Thesis, University of Canterbury. Department of chemisty, 2014. http://hdl.handle.net/10092/10113.
Повний текст джерелаChudziak, Christopher Mark. "Production, characterisation and modification of 1-deoxy-d-xylulose-5-phosphate synthase as a biocatalyst." Thesis, University College London (University of London), 2007. http://discovery.ucl.ac.uk/1445214/.
Повний текст джерелаMichailidou, Freideriki. "Structural and mechanistic studies on the biosynthesis of the 3'-deoxy nucleoside of the pacidamycins." Thesis, University of St Andrews, 2018. http://hdl.handle.net/10023/15668.
Повний текст джерелаHann, Ronald Koy. "Synthesis of N, N'-bis-(3-deoxy-beta-cyclodextrin)-ethyl Endiamine and Potential Photochemical Applications." W&M ScholarWorks, 1987. https://scholarworks.wm.edu/etd/1539625389.
Повний текст джерелаWangpaichitr, Medhi. "The Relevance of mTOR and Hypoxia Inducible Factor to 2-Deoxy-D-Glucose Toxicity in Lung Cancer Cell Lines Under Hypoxia." Scholarly Repository, 2008. http://scholarlyrepository.miami.edu/oa_dissertations/156.
Повний текст джерелаGalvão, Fábio Carrilho. "Caracterização de mutantes condicionais do gene da desoxi-hipusina sintase em Saccharomyces cerevisiae /." Araraquara : [s.n.], 2011. http://hdl.handle.net/11449/87674.
Повний текст джерелаBanca: Nilson Ivo Tonin Zanchin
Banca: Paulo Sergio Rodrigues Coelho
Resumo: O fator de início de tradução 5A (eIF5A) é altamente conservado de arqueas a mamíferos e é essencial para a viabilidade celular. Este fator é a única proteína conhecida que sofre uma modificação pós-traducional única e necessária para a função de eIF5A, em que uma lisina específica é convertida em um resíduo de hipusina pela ação das enzimas desoxi-hipusina sintase (Dys1) e desoxi-hipusina hidroxilase (Lia1). Inicialmente, eIF5A foi relacionada à etapa do início da tradução, porém, dados recentes sugerem a sua atuação na etapa de elongação ao invés de início. No entanto, além do fato de a função específica de eIF5A na célula não ser conhecida, o papel da hipusinação para o funcionamento de eIF5A também não é conhecido. Diante disso, o objetivo deste trabalho é caracterizar mutantes condicionais para o gene da desoxi- hipusina sintase e, dessa forma, contribuir para o entendimento não só da função da hipusinação sobre eIF5A, mas também para o entendimento da função específica de eIF5A na célula. Para isso, foram iniciadas análises de caracterização fenotípica com os alelos dys1Δ1-28 e dys1W75R/T118A/A147T (dys1-1). Inicialmente, foi realizada a subclonagem do alelo dys1Δ1-28 , uma vez que, por ter sido identificado em um rastreamento de duplo-híbrido, este alelo estava em fusão com a região codificadora do domínio de ativação de Gal4. Porém, após realização da subclonagem, ou seja, quando na ausência do domínio de ativação, este alelo não apresentou o fenótipo condicional de crescimento inicialmente observado. Portanto, o mutante se tornou impróprio para a realização dos ensaios subsequentes e foi descartado. Em seguida, foram iniciadas as análises com o alelo dys1-1, nas quais foi observada diminuição nos níveis totais de Dys1 mutada, e consequentemente, diminuição nos níveis de hipusinação. Devido a isso ... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: The translation initiation factor 5A (eIF5A) is highly conserved from archaea to mammals and is essential for cell viability. This factor is the only known protein that undergoes an unique and essential post-translational modification, in which a specific lysine residue is converted into hypusine by the action of the enzymes deoxyhypusine synthase (Dys1) and deoxyhypusine hydroxylase (Lia1). Initially, eIF5A was related to the initiation step of translation, however, recent data suggest a role in the elongation step of translation. However, besides the fact that the specific function of eIF5A in the cell is still obscure, the role of hypusination in eIF5A function is unknown. Thus, the goal of this project is to characterize conditional mutants of the deoxyhypusine synthase gene and thereby contribute to the understanding not only the function of hypusination in eIF5A, but also of the specific role of eIF5A in the cell. We started a phenotypic characterization of two different alleles: dys1Δ1-28 and dys1W75R/T118A/A147T (dys1-1). Initially, we performed a subcloning of the allele dys1Δ1-28 , once the allele was fused with the coding region of GAL4 activation domain, due to the fact that this allele is devived of a two-hybrid screening. However, after performing the subcloning, that is, in the absence of the activation domain, this allele showed no conditional growth phenotype as originally observed. Therefore, this mutant has become improper to carry out the subsequent analysis and was discarted. Then, the analyses with dys1-1 allele were initiated, in which it was observed a decrease in total levels of Dys1 and, consequently, a decrease in the hypusination levels. Because of that, this allele shows a decrease in cell growth rate and growth arrests after 24 hours in medium lacking the osmotic regulator. However, this growth arrest is not followed by cell lysis. Furthermore, the mutant ... (Complete abstract click electronic access below)
Mestre
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Повний текст джерелаScience, Faculty of
Chemistry, Department of
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