Дисертації з теми "Structural binding characterization"
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Hoy, Julie Anne. "Structural characterization of ligand binding in hexacoordinate hemoglobins." [Ames, Iowa : Iowa State University], 2006.
Знайти повний текст джерелаPang, Bo, and 龐博. "Structural characterization of H1N1 nucleoprotein-nucleozin binding sites." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2013. http://hdl.handle.net/10722/205641.
Повний текст джерелаpublished_or_final_version
Physiology
Doctoral
Doctor of Philosophy
Butan, Carmen Crina. "Structural characterization of the RNA binding domain of BTV non-structural protein 2." Thesis, Birkbeck (University of London), 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.414198.
Повний текст джерелаSantelli, Eugenio. "The binding of MEF2A to DNA : biochemical and structural characterization /." [S.l.] : [s.n.], 2000. http://e-collection.ethbib.ethz.ch/show?type=diss&nr=13752.
Повний текст джерелаGilbert, Sunny Deshea. "Biochemical and structural characterization of ligand binding by the purine riboswitch." Connect to online resource, 2007. http://gateway.proquest.com/openurl?url_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&res_dat=xri:pqdiss&rft_dat=xri:pqdiss:3256472.
Повний текст джерелаDesjardins, Geneviève. "Structural characterization of DNA binding and autoinhibition by the Ets1 transcription factor." Thesis, University of British Columbia, 2015. http://hdl.handle.net/2429/52606.
Повний текст джерелаMedicine, Faculty of
Biochemistry and Molecular Biology, Department of
Graduate
Brooksbank, Robert Alan. "Expression, purification and structural characterization of the DNA-binding domain of BZLF1." Thesis, University of Cambridge, 1994. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.321537.
Повний текст джерелаOke, Muse. "Functional and structural characterization of the transferrin binding protein A from Neisseria meningitidis." Thesis, Birkbeck (University of London), 2005. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.413693.
Повний текст джерелаJuntunen, K. (Kari). "Functional and structural characterization of nuclear vitamin D receptor and its ligand binding domain." Doctoral thesis, University of Oulu, 2002. http://urn.fi/urn:isbn:9514268784.
Повний текст джерелаSharif, Azar. "Structural characterization of the polycomb repressor complex 1 binding partner ubiquitin specific protease 11." Thesis, Imperial College London, 2013. http://hdl.handle.net/10044/1/39355.
Повний текст джерелаTaubert, Alexander. "Characterization of DNA binding of the two zinc finger domains of transcription factor zBED6." Thesis, Uppsala universitet, Institutionen för biologisk grundutbildning, 2019. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-396233.
Повний текст джерелаGhosh, Madhumita. "Structural and biochemical characterization of proteins involved in cancer." [S.l. : s.n.], 2005. http://deposit.ddb.de/cgi-bin/dokserv?idn=974284823.
Повний текст джерелаShanker, Sreejesh. "Structural and biochemical characterization of cell cycle regulatory proteins and their inhibitors." [S.l. : s.n.], 2005. http://deposit.ddb.de/cgi-bin/dokserv?idn=974284882.
Повний текст джерелаKumar, Sandeep. "Biochemical, Mechanistic, and Structural Characterization of DNA Polymerase X from African Swine Fever Virus." The Ohio State University, 2008. http://rave.ohiolink.edu/etdc/view?acc_num=osu1211380265.
Повний текст джерелаBohl, Casey Edward. "Structural characterization of androgen receptor interactions with nonsteroidal ligands." Connect to resource, 2005. http://rave.ohiolink.edu/etdc/view?acc%5Fnum=osu1116362600.
Повний текст джерелаBortoluzzi, Alessio. "Structural characterization of Mycobacterium tuberculosis RNA polymerase binding protein A (RbpA) and its interactions with sigma factors." Thesis, University of Leicester, 2013. http://hdl.handle.net/2381/28401.
Повний текст джерелаCollins, Courtney E. "Characterization of SPOC/NCoR Binding: A Thermodynamic and Structural Analysis of Corepressors in the Notch Signaling Pathway." University of Cincinnati / OhioLINK, 2017. http://rave.ohiolink.edu/etdc/view?acc_num=ucin1510926401526291.
Повний текст джерелаWEGRECKI, MARCIN. "Structural, biophysical and functional characterization of Nop7-Erb1-Ytm1 complex and its implications in eukaryotic ribosome biogenesis." Doctoral thesis, Universitat Politècnica de València, 2015. http://hdl.handle.net/10251/55941.
Повний текст джерела[ES] El ensamblaje de ribosomas es uno de los procesos más importantes y costosos energéticamente en una célula eucariota. A pesar de ello, se sabe relativamente poco acerca de la gran mayoría de los eventos y factores implicados en la síntesis de las subunidades ribosomales. La maduración de ribosomas comprende numerosos pasos de procesamiento del rRNA que requieren la asociación y disociación de más de doscientos factores de ensamblaje. Esas proteínas establecen una compleja red de interacciones que son esenciales para que el proceso pueda llevarse a cabo. Los estudios realizados en Saccharomyces cerevisiae han permitido la identificación de algunas correlaciones genéticas y funcionales entre los factores prerribosomales. Es el caso del heterotrímero formado por Nop7, Erb1 e Ytm1 (complejo PeBoW en mamíferos), que es imprescindible para la correcta formación de la subunidad 60S. La ausencia de cualquiera de las tres proteínas es inviable y también se conocen ciertas variantes truncadas que alteran el procesamiento del rRNA 27SA2 y de este modo afectan la proliferación celular. Se ha demostrado que Nop7 y Erb1 se asocian al rRNA y que su reclutamiento al pre60S ocurre antes de la unión a Ytm1. Además se sabe que el trímero tiene que separarse de la partícula prerribosomal emergente con el fin de favorecer su maduración. A pesar de su gran relevancia en la célula, no está claro el papel exacto del complejo PeBoW y tampoco se dispone de conocimientos suficientes acerca de las interacciones intermoleculares que lo mantienen. Durante el desarrollo de este proyecto se ha llevado a cabo un exhaustivo análisis bioquímico y estructural del trímero Nop7-Erb1-Ytm1 procedente de S. cerevisiae y del hongo termofílico Chaetomium thermophilum. En este trabajo hemos sido capaces de reconstituir el complejo estable in vitro que posteriormente se ha utilizado en los ensayos de cristalización, con los que hemos podido resolver la estructura del dominio carboxi-terminal de Erb1 de levadura, cuyo plegamiento corresponde a una hélice enrollada (ß-propeller) de siete hojas. Gracias a la información estructural, hemos demostrado que esa parte de la proteína es capaz de unir RNA in vitro, lo que puede ser una propiedad importante para su función. Además, a pesar de los estudios anteriores que sugerían que la hélice enrollada de Erb1 no era esencial en la biogénesis del ribosoma, hemos resuelto la estructura cristalina de la proteína Ytm1 unida al dominio C-terminal de Erb1 de C. thermophilum. Ese descubrimiento nos ha permitido redefinir las interacciones macromoleculares que mantienen el complejo. Inicialmente hemos confirmado que el extremo amino-terminal de Nop7 interacciona con Erb1. A continuación, hemos demostrado que el dominio WD40 de Ytm1 se une al ß-propeller de Erb1 con una buena afinidad. Después de un detallado análisis de la superficie involucrada en la formación del dímero, hemos sido capaces de diseñar una variante mutada de Erb1 que se asocia más débilmente con Ytm1. Los hallazgos estructurales y biofísicos se han confirmado in vivo usando S. cerevisiae donde hemos demostrado que una mutación puntual que disminuye la afinidad de unión entre los dominios C-terminales de Erb1 e Ytm1 manifiesta un efecto negativo sobre el crecimiento de levadura porque interfiere con la síntesis de 60S. Nuestros resultados establecen un buen ejemplo de una superficie conservada involucrada en interacciones proteína-proteína, que podría considerarse una buena diana para inhibir la proliferación celular eucariota.
[CAT] L'ensamblatge de ribosomes és un dels processos més importants i energèticament costosos en una cèl·lula eucariota. Tot i això, es coneix relativament poc de la majoria dels factors implicats en la síntesi de les subunitats ribosomals. La maduració de ribosomes compren moltes etapes de processament del rRNA que requereix l'associació i dissociació de més de dos-cents factors d'ensamblatge. Aquestes proteïnes estableixen una complexa xarxa de interaccions que són essencials perquè el procés es pugi dur a terme. Els estudis realitzats en Saccharomyces cerevisiae han permès la identificació de algunes correlacions genètiques i funcionals entre els factors pre-ribosomals. Aquest és el cas del heterotrímer comprés per Nop7, Erb1 i Ytm1 (complex PeBoW en mamífers), que és imprescindible per a la correcta formació de la subunitat 60S. L'absència de qualsevol de les tres proteïnes és inviable i també és coneixen certes variants truncades que alteren el processament del rRNA 27SA3 i que d'aquesta manera afecten a la proliferació cel·lular. S'ha demostrat que Nop7 i Erb1 s'associen al rRNA i que el seu reclutament al pre60S té lloc abans de l'unió a Ytm1. A més a més, es sap que el trímer ha de separar-se de la partícula pre-ribosomal emergent per tal que es produeixi la seua maduració. Malgrat la seua rellevància en la cèl·lula, no s'ha aclarit el paper exacte del complex PeBoW i tampoc n'hi ha coneixements suficients de les interaccions intermoleculars que el mantenen. Durant el desenvolupament d'aquest projecte s'ha dut a terme un exhaustiu anàlisi bioquímic i estructural del trímer Nop7-Erb1-Ytm1 de S. cerevisiae i del fong termofílic Chaetomium thermophilum. En aquest treball hem estat capaços de reconstituir el complex estable in vitro que posteriorment s'ha utilitzat en el assajos de cristal·lització, amb els que hem pogut resoldre l'estructura del domini carboxi-terminal de Erb1 de llevat i que té un plegament corresponent a una hèlix enrotllada (ß-propeller) de set fulles. Gràcies a la informació estructural, hem pogut demostrar que aquesta part de la proteïna té la capacitat d'unir RNA in vitro, el que pot ser una propietat important per a la seua funció. A més a més, malgrat que els estudis anteriors suggerien que la hèlix enrotllada de Erb1 no era essencial en la biogènesis del ribosoma, hem pogut resoldre la estructura cristal·lina de la proteïna Ytm1 unida al domini C-terminal de Erb1 de C. thermophilum. Aquest descobriment ens ha permès redefinir les interaccions macromoleculars que mantenen el complex. Inicialment, hem confirmat que l'extrem amino-terminal de Nop7 interacciona amb Erb1. A continuació, hem demostrat que el domini WD40 de Ytm1 s'uneix al ß-propeller de Erb1 amb bona afinitat. Després d'un anàlisi detallat de la superfície involucrada en la formació del dímer, hem estat capaços de dissenyar una variant mutada de Erb1 que s'associa més dèbilment amb Ytm1. Les dades estructurals i biofísiques s'han confirmat in vivo utilitzant S. cerevisiae on hem demostrat que una mutació puntual que disminueix l'afinitat d'unió entre els dominis C-terminals de Erb1 i Ytm1 manifesta un efecte negatiu en el creixement del llevat perquè interfereix amb la síntesi del 60S. Els nostres resultats estableixen un bon exemple de una superfície conservada involucrada en interaccions proteïna-proteïna, que es podria considerar una bona diana per a inhibir la proliferació cel·lular eucariota.
Wegrecki, M. (2015). Structural, biophysical and functional characterization of Nop7-Erb1-Ytm1 complex and its implications in eukaryotic ribosome biogenesis [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/55941
TESIS
Blissing, Annica. "Thiopurine S-methyltransferase - characterization of variants and ligand binding." Licentiate thesis, Linköpings universitet, Kemi, 2017. http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-136558.
Повний текст джерелаValiveti, Aswani Kumar. "Structural characterization of metal and DNA binding to DREAM protein, a calcium sensing transcriptional repressor in pain modulation." College Park, Md. : University of Maryland, 2006. http://hdl.handle.net/1903/3975.
Повний текст джерелаThesis research directed by: Molecular and Cell Biology. Title from t.p. of PDF. Includes bibliographical references. Published by UMI Dissertation Services, Ann Arbor, Mich. Also available in paper.
Manoharan, Malini. "Genomic, structural and functional characterization of odorant binding proteins in olfaction of mosquitoes involved in infectious disease transmission." Phd thesis, Université de la Réunion, 2011. http://tel.archives-ouvertes.fr/tel-00979587.
Повний текст джерелаGrimm, Nicole Elena. "Characterization of the Schizosaccharomyces pombe protection of telomeres 1 (Pot1) DNA-binding domains by biochemical and structural techniques." Connect to online resource, 2008. http://gateway.proquest.com/openurl?url_ver=Z39.88-2004&rft_val_fmt=info:ofi/fmt:kev:mtx:dissertation&res_dat=xri:pqdiss&rft_dat=xri:pqdiss:1460859.
Повний текст джерелаOkamoto, Patricia Michiyo. "Nitrate reductase of Neurospora crassa : characterization of its gene, nit-3, and structural studies of its heme-binding domain /." The Ohio State University, 1992. http://rave.ohiolink.edu/etdc/view?acc_num=osu1487776801320659.
Повний текст джерелаHébert-Losier, Andréa 1983. "Structural and functional characterization of a novel endogenous steroid, estradienolone (ED), in human pregnancy." Thesis, McGill University, 2008. http://digitool.Library.McGill.CA:80/R/?func=dbin-jump-full&object_id=116111.
Повний текст джерелаJakob, Leonhard [Verfasser], and Gunter [Akademischer Betreuer] Meister. "Structural and functional characterization of the RNA-binding proteins Loquacious and Brain tumor from Drosophila melanogaster / Leonhard Jakob ; Betreuer: Gunter Meister." Regensburg : Universitätsbibliothek Regensburg, 2017. http://d-nb.info/1129956628/34.
Повний текст джерелаNickolaus, Chen [Verfasser], and Wolfgang E. [Akademischer Betreuer] Trommer. "The Molten Globule State of Maltose-Binding Protein: Structural Characterization by Electron Paramagnetic Resonance Spectroscopy / Chen Nickolaus ; Betreuer: Wolfgang E. Trommer." Kaiserslautern : Technische Universität Kaiserslautern, 2017. http://d-nb.info/1123572135/34.
Повний текст джерелаSturm, Noé. "Characterization of natural product biological imprints for computer-aided drug design applications." Thesis, Strasbourg, 2015. http://www.theses.fr/2015STRAF059/document.
Повний текст джерелаCan computational binding site similarity tools verify the hypothesis: “Biosynthetic moldings give potent biological activities to natural products”? To answer this question, we designed a tool modeling binding site properties according to solvent exposure. The method showed interesting characteristics but suffers from sensitivity to atomic coordinates. However, existing methods have delivered evidence that the hypothesis was valid for the flavonoid chemical class. In order to extend the study, we designed an automated pipeline capable of searching natural products biosynthetic enzyme structures embedding ligandable catalytic sites. We collected structures of 117 biosynthetic enzymes. Finally, according to structural investigations of biosynthetic enzymes, we characterized diverse substrate-enzyme binding-modes, suggesting that natural product biological imprints usually do not agree with the “key-lock” model
Sitar, Tomasz. "Structural basis for the inhibition of insulin-like growth factors by insulin-like growth factor-binding proteins and structural and biochemical characterization of formins - the actin nucleating factors." kostenfrei, 2007. http://mediatum2.ub.tum.de/doc/652583/652583.pdf.
Повний текст джерелаFleming, Christopher Daniel Redinbo Matthew Robert. "Structural insights into xenobiotic and organophosphate binding by human carboxylesterase 1 and efforts made towards the characterization of the androgen receptor modulator MAGE-11." Chapel Hill, N.C. : University of North Carolina at Chapel Hill, 2007. http://dc.lib.unc.edu/u?/etd,1228.
Повний текст джерелаTitle from electronic title page (viewed Mar. 26, 2008). "... in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the Department of Biochemistry and Biophysics." Discipline: Biochemistry and Biophysics; Department/School: Medicine.
Lange, Anja. "Structural characterization of the interaction of the Stam2's ubiquitin binding domains with ubiquitin chains by NMR : Cooperativity or not, that is the question !" Thesis, Lyon 1, 2010. http://www.theses.fr/2010LYO10308.
Повний текст джерелаFrom the discovery of ubiquitin and its function as signal for proteasomal degradation over 20 years ago to this days, it became evident that ubiquitin is a universal signal in eukaryotic cells. Ubiquitin in its different forms is involved in many versatile cellular processes. Knowing that the ubiquitin signal is differently translated, depending on its occurrences as mono-ubiquitin or poly-ubiquitin, raises the question: how do cells distinguish between the different occurrences of ubiquitin and translate it into the proper response? Proteins interacting with ubiquitin contain so called ubiquitin binding domains (UBDs), whereas the affinities to ubiquitin vary from a few _M to mM. So far only three (K63, K48 and linear chains) out of the eight possible chain-linkages can be produced in sufficient amounts to characterize their interaction with UBDs. K48- and K63- linked ubiquitin chains regulate different cellular events and need to be recognized by different proteins. Thus, it is of prime importance to characterize the binding of different UBDs to these two kinds of ubiquitin chains, as it can give important clues related to the general mechanism of chain discrimination by ubiquitin adapter proteins. Some isolated UBDs exhibit a preference for one chain linkage type over the other, whereas others do not discriminate between mono-ubiquitin or K63- and K48-linked chains. Interestingly, many ubiquitin adapter proteins harbor more than one UBD. STAM2 is a ubiquitin adapter protein, that is involved in endosomal receptor sorting and supposed to preferentially bind mono-ubiquitin and K63- over K48-linked ubiquitin. STAM2 contains two UBDs (a VHS and UIM domain) that were shown to bind to ubiquitin . The current manuscript shows that STAM2’s SH3 domain binds ubiquitin as well. To understand the function of the sequential arrangement of three UBDs in one protein, first binding of the individual VHS and UIM domains to monoubiquitin as well as K48- and K63-linked di-ubiquitin was investigated. This work shows, that the VHS domain displays a different mode of binding for K63- and K48-linked diubiquitin. In spite of the fact, that the apparent Kd for both chains is the same, only one VHS domain can bind to K48-linked di-ubiquitin chains (with a preference for the distal domain), whereas K63-linked di-ubiquitin can accommodate two VHS domains at a time. Since no conclusion can be drawn with respect to the apparent Kds, the different binding modes might gain more impact in consideration of the ensemble of three UBDs. Results presented in this manuscript, based on a construct containing the VHS and UIM domain, show that binding to K63- but not K48-linked di-ubiquitin is cooperative
Wang, Qianmin Verfasser], and Elena [Akademischer Betreuer] [Conti. "Structural and Biochemical Characterization of Cell Shaping Proteins : 1. Microtubule Binding Protein p150glued and 2. Intraflagellar Transport Protein 172 / Qianmin Wang ; Betreuer: Elena Conti." München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2017. http://d-nb.info/1148276807/34.
Повний текст джерелаTavares, Macedo Joana [Verfasser], and Bärbel [Akademischer Betreuer] Blaum. "Production and glycan binding characterization of human properdin and structural elucidation of c-Jun N-terminal kinase 3 inhibitors / Joana Tavares Macedo ; Betreuer: Bärbel Blaum." Tübingen : Universitätsbibliothek Tübingen, 2019. http://d-nb.info/1201644925/34.
Повний текст джерелаRechlin, Chris [Verfasser], and Gerhard [Akademischer Betreuer] Klebe. "Insights into Protein-Ligand Molecular Recognition: Thermodynamic, Kinetic and Structural Characterization of Inhibitor Binding to Aldose Reductase and Carbonic Anhydrase II / Chris Rechlin ; Betreuer: Gerhard Klebe." Marburg : Philipps-Universität Marburg, 2016. http://d-nb.info/1119318017/34.
Повний текст джерелаPort, Sarah A. [Verfasser], Ralph H. [Akademischer Betreuer] Kehlenbach, Achim [Akademischer Betreuer] Dickmanns, and Heinz [Akademischer Betreuer] Neumann. "Structural and Functional Characterization of CRM1-Nup214 Interactions Reveals Multiple FG-binding Sites Involved in Nucleocytoplasmic Transport / Sarah A. Port. Gutachter: Achim Dickmanns ; Heinz Neumann. Betreuer: Ralph H. Kehlenbach." Göttingen : Niedersächsische Staats- und Universitätsbibliothek Göttingen, 2015. http://d-nb.info/1076398685/34.
Повний текст джерелаGallego, Alonso Pablo. "Structural studies on protein-protein interactions: Analysis of the regulation of the DYNLL/LC8 binding to Nek9 and characterization of the enzymes composing the arginine deiminase pathway in mycoplasma penetrans." Doctoral thesis, Universitat Autònoma de Barcelona, 2014. http://hdl.handle.net/10803/285569.
Повний текст джерелаProtein-Protein Interactions (PPIs) are intentional physical contacts established between two or more proteins. These interactions form the large protein interaction network and are the core of the entire interatomic system of any living cell. Flaws in the interaction network by aberrant PPIs involve signal transduction fails, protein aggregation and a completely loss of cell's regulation. Indeed, missense PPIs are the basis of multiple diseases, such as Alzheimer's disease and cancer. The significance of the interatomic system makes worth to studying PPIs, their functions and structural properties, in different models. In the present work X-ray crystallography is the main technique used, in addition to other methods to study Protein-Protein interactions. The study of protein-protein interactions is performed through two different experimental models: In the first model, we analyzed the role of phosphorylation in the interaction of a peptide derived from the mitotic kinase Nek9 with its binding partner LC8; In the second model, we decipher the structures of the three enzymes composing the arginine deiminase pathway in Mycoplasma penetrans. The NIMA family protein kinase Nek9/Nercc1 plays a main role in the control of the mitotic spindle. DYNLL/LC8 was originally described as a component of the dynein complex, but the recent discovery of multiple interaction partners for LC8 has proposed that it has a general role as a dimerization hub that organizes different protein partners. Recent experiments suggested that LC8 binding to Nek9 was regulated by Nek9 autophosphorylation. The present work sheds light into a novel phosphorylation regulatory mechanism that interferes with LC8 protein-protein complex formation. The metabolism of arginine towards ATP synthesis has been considered a major source of energy for microorganisms such as Mycoplasma penetrans in anaerobic conditions. Additionally, this pathway has also been implicated in pathogenic and virulence mechanism of certain microorganisms, i.e. protection from acidic stress during infection. In this work we present the crystal structures of the three enzymes composing the gene cluster of the arginine deiminase pathway from M.penetrans: arginine deiminase (ADI), ornithine carbamoyltransferase (OTC) and carbamate kinase (CK). M.penetrans ADI and CK structures disclose the structural conformation shifts upon the reaction mechanism of these proteins. In the case of M.penetrans OCT its dodecameric quaternary structure is compared with other organisms (including some thermophiles), revealing the formation common quaternary structure arranged interfaces with a low sequence homology. The results collected in this thesis emphasize the important contribution of X-ray crystallography for studding PPIs properties. PPIs regulation, structural and functional differences depending on their association, posttranslational modifications and the environment adaptation thought evolution are some of the PPI properties and dependence factors studied in this thesis work.
Kowalska, Kaja [Verfasser], Tad A. [Akademischer Betreuer] Holak, Bernd [Akademischer Betreuer] Reif, and Robert [Akademischer Betreuer] Huber. "Biochemical and biophysical characterization of CD44 and its binding partner, hyaluronic acid and structural investigations of the ubiquitin-like protein 5 / Kaja Kowalska. Gutachter: Bernd Reif ; Robert Huber. Betreuer: Tad A. Holak." München : Universitätsbibliothek der TU München, 2012. http://d-nb.info/1031513604/34.
Повний текст джерелаBatista, Adelina Braga. "CaracterizaÃÃo estrutural da Mo-CBP3, uma albumina 2S de sementes de Moringa oleifera lamarck e seu modo de aÃÃo contra fungos fitopatogÃnicos." Universidade Federal do CearÃ, 2013. http://www.teses.ufc.br/tde_busca/arquivo.php?codArquivo=10389.
Повний текст джерелаMo-CBP3 à uma proteÃna ligante à quitina, purificada de sementes de Moringa oleifera, com amplo espectro de aÃÃo contra fungos fitopatogÃnicos. No presente trabalho, novas propriedades estruturais da Mo-CBP3 sÃo descritas, revelando correlaÃÃo entre sua estabilidade estrutural e atividade antifÃngica. Em adiÃÃo, para melhor compreensÃo dos mecanismos pelos quais essa proteÃna exerce aÃÃo antifÃngica, sua habilidade de induzir a produÃÃo endÃgena de espÃcies reativas de oxigÃnio e de causar alteraÃÃes morfolÃgicas e ultraestruturais foi analisada, usando Fusarium solani como modelo. F. solani à uma espÃcie de fÃcil manuseio e desenvolvimento rÃpido, ideal para ensaios in vitro, e de relevÃncia, por se tratar de um fungo que ataca culturas economicamente importantes. Com foco na utilizaÃÃo segura da Mo-CBP3 como agente quÃmico contra fungos, seus efeitos citotÃxicos sobre cÃlulas eucariÃticas tambÃm foram investigados. Mo-CBP3 à uma proteÃna ligante à quitina de 18,0 kDa, de acordo com PAGE-SDS. Todavia, anÃlise por espectrometria de massas revelou que essa proteÃna consiste de mÃltiplas isoformas com massas moleculares variando entre 12,2 e 12,3 kDa. Mo-CBP3 à composta por duas cadeias polipeptÃdicas de 5,0 e 9,0 kDa, denominadas de cadeia A e cadeia B, respectivamente. A cadeia B contÃm a sequÃncia NH2-terminal representada por CPAIQRCCQQLRNIQPPCRCCQ, enquanto que a cadeia A tem o resÃduo NH2-terminal bloqueado. cDNA codificador da cadeia B foi obtido com iniciadores sintetizados a partir de sua sequÃncia NH2-terminal. AnÃlises in silico das sequÃncias de nucleotÃdeos e de aminoÃcidos deduzida confirmaram a presenÃa de isoformas e massa molecular da Mo-CBP3 e identificaram sÃtios potenciais de O-glicosilaÃÃo e fosforilaÃÃo. AlÃm disso, similaridades entre Mo-CBP3 e outras proteÃnas de M. oleifera, bem como com albuminas 2S, foram detectadas. A estrutura secundaria da Mo-CBP3 à composta por 30,3% α-hÃlices, 16,3% folhas β, 22,3% voltas e 30,4% estruturas ao acaso. Na espectroscopia de fluorescÃncia, excitaÃÃes de uma soluÃÃo da Mo-CBP3 a 280 nm e 295 nm produziram emissÃo mÃxima a 303 e 309 nm, respectivamente. A estrutura da Mo-CBP3 à altamente estÃvel, se apresentando indiferente Ãs mudanÃas de temperatura e pH. Mo-CBP3 (0,05-0,1 mg/mL) se mostrou capaz de inibir a germinaÃÃo de conÃdios de vÃrios fungos fitopatogÃnicos, incluindo F. solani, F. oxysporum, Colletotrichum musae e C. gloeosporioides. Similarmente, Mo-CBP3 (0,05 mg/mL) foi capaz de inibir o crescimento micelial de F. solani e apresentou tanto efeito fungistÃtico como fungicida, dependendo da concentraÃÃo usada. LigaÃÃo da Mo-CBP3 à superfÃcie de cÃlulas fÃngicas ocorre, pelo menos em parte, via interaÃÃo eletrostÃtica, jà que NaCl 0,15 M aboliu seu efeito inibitÃrio. Mo-CBP3 induziu a produÃÃo de espÃcies reativas de oxigÃnio e causou perda de assimetria e deformaÃÃes em cÃlulas de F. solani. DesorganizaÃÃo do sistema de endomembranas, condensaÃÃo do citosol e aumento de vacuolizaÃÃo tambÃm foram observados. Mo-CBP3 nÃo mostrou atividade hemolÃtica e nem foi capaz de alterar a viabilidade das cÃlulas MCF-7 e Caco-2, sugerindo que essa proteÃna nÃo à tÃxica para cÃlulas humanas. Com base na alta estabilidade e no amplo espectro de aÃÃo contra fungos fitopatogÃnicos em baixas concentraÃÃes e, tambÃm, na ausÃncia de citotoxicidade para cÃlulas humanas testadas, Mo-CBP3 tem grande potencial para desenvolvimento de novas drogas antifÃngicas ou na produÃÃo de plantas transgÃnicas mais resistentes a fungos.
Mo-CBP3 is a chitin-binding protein purified from Moringa oleifera seeds that displays broad inhibitory activity against phytopathogenic fungi. In this work, we report new structural features of Mo-CBP3 that reveal a correlation between its structural stability and antifungal activity. In addition, to gain better insights into the mechanisms by which this protein acts as an antifungal agent, its ability to induce the endogenous production of reactive oxygen species and to trigger morphologic and ultrastructural alterations were analysed using Fusarium solani as a model. F. solani is an easy-to-handle and fast-developing species, making it ideal for in vitro assays, and it holds relevance as a phytopathogenic fungus that attacks economically important crop plants. To fully explore the biosafety of Mo-CBP3 as a chemical agent against fungi, its cytotoxic effects on eukaryotic cells were also investigated. Mo-CBP3 is a chitin-binding protein of 18.0 kDa, according to SDS-PAGE. However, by mass spectrometry analysis, it was observed that this protein consists of multiple isoforms with molecular masses ranging between 12.2 and 12.3 kDa. Mo-CBP3 is composed by two polypeptide chains of 5.0 and 9.0 kDa, named A and B chain, respectively. The B chain contains the following NH2-terminal sequence CPAIQRCCQQLRNIQPPCRCCQ while the A chain has a blocked NH2-terminal residue. cDNA encoding the B chain was obtained with primers of its NH2-terminal sequence. In silico analyses of nucleotide and deduced amino acid sequences confirmed the presence of isoforms and molecular mass of Mo-CBP3 and identified potential sites of O-glicosylation and phosphorilation. Moreover, similarities between Mo-CBP3 and other M. oleifera proteins as well as 2S albumins were detected. The secondary structure of Mo-CBP3 showed 30.3% α-helices, 16.3% β-sheets, 22.3% turns and 30.4% unordered forms. In the fluorescence spectroscopy, excitation of Mo-CBP3 solution at 280 nm and 295 nm gave emission maxima at 303 and 309 nm, respectively. The Mo-CBP3 structure is highly stable and retains its antifungal activity regardless of temperature and pH. Mo-CBP3 (0.05-0.1 mg/mL) was able to inhibit the conidia germination of several phytopathogenic fungi, including F. solani, F. oxysporum, Colletotrichum musae and C. gloeosporioides. Similarly, Mo-CBP3 was inhibitory to the mycelial mass development of F. solani at 0.05 mg/mL and has both fungistatic and fungicidal effects, depending on the concentration used. Binding of Mo-CBP3 to the fungal cell surface is achieved, at least in part, via electrostatic interactions, as 150 mM NaCl abolished its inhibitory effect. Mo-CBP3 induced the production of reactive oxygen species and caused in F. solani cells a marked loss of asymmetry, deformations and deep wrinkles in comparison to control cells. Disorganisation of the endomembrane system and condensation and shrinkage of cytosol with increased vacuolation and the loss of normal structure and content were also observed. Mo-CBP3 did not show haemolytic activity and it was not capable to alter de viability of both MCF-7 and Caco-2 cells, suggesting that this protein is not toxic for human cells. Based on its high stability and broad-spectrum efficacy against important phytopathogenic fungi at low inhibitory concentrations and absence of cytotoxicity to human cells, Mo-CBP3 has great potential in the development of new antifungal drugs or in transgenic crops with enhanced resistance to fungi.
Betz, Christine [Verfasser], and Oliver [Akademischer Betreuer] Einsle. "Structural characterization of the metal-binding ligands S100A8/S100A9 and S100B of the receptor for advanced glycation end products = Strukturelle Charakterisierung der metallbindenden Liganden S100A8/S100A9 und S100B des Rezeptors für Advanced Glycation End Products." Freiburg : Universität, 2013. http://d-nb.info/1115813455/34.
Повний текст джерелаElison, Kalman Grim. "Purification, functional characterization and crystallization of the PerR peroxide sensor from Saccharopolyspora erythraea." Thesis, Uppsala universitet, Strukturbiologi, 2019. http://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-387943.
Повний текст джерелаEl, Masri Rana. "Remodeling of heparan sulfate : functional and structural characterization of human endosulfatase HSulf-2 The sweet side of extracellular sulfatases Expression and purification of recombinant extracellular sulfatase HSulf-2 allows deciphering of enzyme sub-domain coordinated role for the binding and 6-O-desulfation of heparan sulfate." Thesis, Université Grenoble Alpes (ComUE), 2019. http://www.theses.fr/2019GREAV037.
Повний текст джерелаHeparan Sulfate (HS) are complex polysaccharides involved in many biological processes. The structure of HS is regulated at the cell surface by unique extracellular endosulfatases, the Sulfs. Sulfs dramatically change HS functional properties, thereby being implicated in many physiopathological processes including cancer. Sulfs features two domains: a catalytic domain (CAT) that comprises the active site, and an hydrophilic basic domain (HD) responsible for HS binding. The aim of my PhD project is to characterize the structural and the functional properties of the human for HSulf-2, which remains poorly understood. In this context, we have first studied the enzyme/substrate recognition mechanisms. We identified two novel HS binding motifs on these enzymes implicated in their activity. In addition, using natural and synthetic oligosaccharides, we demonstrated that the HD is not essential for HS recognition, but is directs the processive and orientated desulfation of the polysaccharide. Moreover, we showed that a tetrasaccharide is the minimal oligosaccharide size required for HSulf-2 activity. Our results enabled us to propose a new model depicting the desulfation process of HS by the Sulfs. Second, we have shown that HSulf-2 is a proteoglycan, given that it harbors a unique PTM (Chondroitin Sulfate, CS chain) on its HD domain. This chain decreases enzyme activity and HS binding in vitro. In the tumoral microenvironment, using a murine orthotropic mammary tumor model, we showed that the CS chain is lost by proteolytic processing, leading to the activation of HSulf-2, and the promotion of tumor growth, vascularization and metastasis. Finally, we have undertaken the structural characterization of the Sulfs. For this, we decided to study separately the two domains found in these enzymes (CAT and HD). Crystallogenesis assays were undertaken for the CAT domain to solve its structure by X-ray crystallography, but were unsuccessful. Regarding the HD, we set up a protocol of production and purification of recombinant HD and we initiated NMR studies and other biophysics analyses in order to structurally characterize the domain and to identify the HS binding sites. Our preliminary results suggest that the HD is an unstructured domain, except for its N- and C-terminal parts. Overall, our data provide significant insights into this critical regulatory step of HS function
Faris, Jonathan Scott. "Characterization of the DNA binding properties of the thyroid hormone receptor." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1996. http://www.collectionscanada.ca/obj/s4/f2/dsk3/ftp04/nq21931.pdf.
Повний текст джерелаBaumgärtel, Thomas. "Binding and characterization of fluorescent nano-aggregates on structured surfaces." Doctoral thesis, Universitätsbibliothek Chemnitz, 2012. http://nbn-resolving.de/urn:nbn:de:bsz:ch1-qucosa-91552.
Повний текст джерелаSun, Xueguang. "Characterization of E. coli HFQ structure and its RNA binding properties." Available online, Georgia Institute of Technology, 2006, 2006. http://etd.gatech.edu/theses/available/etd-12052005-145342/.
Повний текст джерелаWartell Roger, Committee Chair ; Chernoff Yury, Committee Member ; Harvey Stephen, Committee Member ; Spiro Stephen, Committee Member ; Williams Loren, Committee Member.
Sun, Xueguang. "Characterization of E coli Hfq structure and its RNA binding properties." Diss., Georgia Institute of Technology, 2005. http://hdl.handle.net/1853/10416.
Повний текст джерелаCarrick, Francine Ellen. "Characterization of bovine insulin like growth factor binding protein-2 : structure and function." Title page, abstract and contents only, 2001. http://web4.library.adelaide.edu.au/theses/09PH/09phc3158.pdf.
Повний текст джерелаChinenov, Yurii. "Molecular characterization of transcription factor GABP : redox regulation, promoter structure, and mechanisms of assembly /." free to MU campus, to others for purchase, 1999. http://wwwlib.umi.com/cr/mo/fullcit?p9962509.
Повний текст джерелаArbuckle, Janeen Lynnae. "Identification and characterization of domains in non-core RAG1." Oklahoma City : [s.n.], 2007.
Знайти повний текст джерелаKiesler, Eva. "Isolation and functional characterization of Hrp65-binding proteins in Chironomus tentans." Doctoral thesis, Stockholm : Institutionen för molekylärbiologi och funktionsgenomik, Univ, 2004. http://urn.kb.se/resolve?urn=urn:nbn:se:su:diva-218.
Повний текст джерелаAlves, Carolina Alpalhão Mantero de Mendonça. "Characterization of the hepatitis delta virus small antigen: intracellular localization, structure, multimerization and RNA binding ability." Doctoral thesis, Instituto de Higiene e Medicina Tropical, 2013. http://hdl.handle.net/10362/19280.
Повний текст джерелаHepatitis delta virus (HDV) is the causative agent of one of the most severe forms of viral hepatitis. It has a small single-stranded circular RNA genome of negative polarity and only one viral protein, the small delta antigen (S-HDAg). Following site-specific RNA editing, a second longer protein is translated, the large delta antigen (L-HDAg). Although these viral proteins share most of their sequence they play distinct roles. S-HDAg is essential for the accumulation of HDV RNAs whereas L-HDAg inhibits HDV replication and is necessary for viral assembly. With such a limited coding capacity HDV must rely extensively on host cell components to complete its replication cycle. The host DNA-directed RNA polymerase II (pol II) is thought to be re-directed to transcribe HDV RNAs. The objective of this study was to further characterize S-HDAg and clarify its role(s) during the HDV replication cycle. We observed that when S-HDAg was expressed in vivo along with replicating HDV RNA it co-located with host pol II. However, such co-localization was also observed in the presence of non-replicating HDV RNAs or when replication was inhibited by specific doses of -amanitin. Thus, we propose that S-HDAg is essential for HDV RNA accumulation by stabilizing or protecting the viral RNAs rather than acting as a direct player in HDV RNA transcription. Additionally, we observed that S-HDAg located in nucleolus when expressed in the absence of HDV RNA, and co-located with host nucleolin. However, in the presence of non-replicating HDV RNAs, S-HDAg moved to the nucleoplasm whereas nucleolin was unchanged. This suggests that S-HDAg is not interacting directly with nucleolin. In our examination of S-HDAg‟s structural features we applied a meta-predictor of intrinsic disorder, PONDR-FIT. It predicted that full-length S-HDAg has extensive intrinsic disorder. This result was confirmed in vitro by circular dichroism measurements that indicated no more than 30% of S-HDAg amino acids adopted an -helical structure. Such a lack of a well-defined rigid structure is expected to grant flexibility to the antigen allowing it to interact with several partners and perform distinct roles during the HDV replication cycle. Protein multimerization was studied by dynamic light scattering. Data analysis indicated that purified recombinant S-HDAg was able to assemble into homomultimers as high as dodecamers. Similarly, denaturing polyacrylamide gel electrophoresis with prior cross-linking indicated formation of at least hexamers and octamers. Similar multimers were observed for S-HDAg present in virus-like particles indicating that S-HDAg multimerization also occurs in vivo.Finally we examined the ability of S-HDAg to bind nucleic acids in vitro. Both multimers and monomers bound to conformations of both RNA and DNA. Such a lack of specificity was probably due to electrostatic interactions between the positively-charged S-HDAg (+12) and negatively-charged nucleic acids. We propose that in vivo, extensive post-translational phosphorylation of S-HDAg reduces the positive charge, thereby contributing to interactions more specific for HDV RNAs and possibly dependent upon protein multimerization. Despite our observations presented here, some issues relating to our aims remain unresolved.
Mariotti, Paolo <1980>. "Characterization of the structure and iron uptake mechanisms of the Staphylococcus aureus ferric hydroxamate-binding lipoprotein FhuD2." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2012. http://amsdottorato.unibo.it/4821/.
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