Дисертації з теми "Prone proteins"
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Malm, Linus. "Size determination of hyaluronan and multivariate analysis of amyloid prone proteins." Doctoral thesis, Umeå universitet, Institutionen för medicinsk kemi och biofysik, 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:umu:diva-46601.
Повний текст джерелаBerger, Z. "The biology of aggregate-prone proteins and possible therapeutic interventions against their toxicity." Thesis, University of Cambridge, 2006. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.596587.
Повний текст джерелаGallo, Annastassia Dawn. "Homeostasis and trafficking of hydrolysis-prone metals in cells, proteins, and small molecules." Diss., Temple University Libraries, 2019. http://cdm16002.contentdm.oclc.org/cdm/ref/collection/p245801coll10/id/568230.
Повний текст джерелаPh.D.
Nature uses inorganic elements for biological processes based on the useful chemistry, abundance, and availability of each metal. Transition metals are critical in the biogeochemical cycling of essential elements and the bioinorganic chemistry of organisms. Hydrolysis-prone metals such as iron and titanium are abundant on Earth but are mostly insoluble in oxic aqueous environments. Nearly every organism requires iron for survival, therefore Nature evolved to stabilize iron from hydrolysis and hydrolytic precipitation through protein and small molecule mechanisms. Like iron, titanium primarily exists as insoluble mineral oxides and is second only to iron as the most abundant transition metal in the Earth’s crust. Despite the reputation as an inert and insoluble metal, titanium can be solubilized and made bioavailable through by chemical and biological weathering. Currently there is no known native role for titanium, however it is quite bioactive. As a stronger Lewis acid, titanium can compete with iron in binding to biomolecules and proteins. It is of interest to investigate the interactions between hydrolysis-prone metals and biological systems, from whole cell organisms to proteins and small molecules. The non-pathogenic bacterium Rhodococcus ruber GIN-1 was isolated for its ability to strongly adhere to titanium dioxide (TiO2) over other metal oxides, providing an opportunity to study the interactions between whole bacterial cells and metal oxides. The GIN-1 strain incorporates Ti(IV) ions into its biomass after adherence to anatase, rutile, and a mixture of the two morphologies. Six metals were quantitated in TiO2-exposed and control (unexposed) cells by inductively coupled plasma optical emission spectroscopy. The exposure to TiO2 caused a significant uptake of titanium with concomitant loss of iron, zinc, and possibly manganese. A collaborative project with the Strongin laboratory at Temple University works to develop stable, biomaterial photocatalysts for environment remediation of toxic inorganic contaminants. Ferritins are a class of proteins that mineralizes and stores iron as a non- toxic ferrihydrite nanoparticle. These proteins can be photoactivated with ultraviolet light to release iron from its core to remediate environmental contaminants. Ferritin can be sensitized with plasmonic gold nanoparticles to extend the photoactivity of the catalyst to the visible spectrum. Work in this thesis highlights the contribution to this collaboration from the Valentine laboratory, included the expression and purification of proteins in E. coli (human H-chain ferritin, human L-chain ferritin, and bacterial DNA protection from starved cells protein), mutation of proteins to improve sensitization of catalyst, and biomineralization with iron and titanium. The trafficking of hydrolysis prone metals is vital for the survival of nearly every organism. Iron transport proteins such as transferrins are studied to understand how nature utilizes a difficult essential metal across the domains of life. Most transferrins have two homologous lobes and are believed to have evolved from a gene duplication of a monolobal transferrin. The ascidian Ciona intestinalis has genes for both a bilobal and monolobal transferrin. Nicatransferrin (nicaTf), the monolobal transferrin from C. intestinalis, is a primitive protein that may provide insight on the evolution of transferrins in higher organisms. It is advantageous to use E. coli expression systems to produce recombinant proteins, however protein misfolding and aggregation can be a concern. To improve expression of nicaTf in E. coli, codon optimization and disulfide bonded protein expression were used. Finally, siderophores are small, high affinity iron-chelating molecules secreted from lower organisms that scavenge iron in iron-limiting conditions. R. ruber GIN-1 and R. ruber DSM 43338 strains both secrete siderophores in artificial seawater media. There are several siderophores identified from Rhodococcus species, however none have been reported from any R. ruber strain. A new siderophore was isolated and preliminary work has been done to purify and characterize the molecule. Understanding the siderophore- metal ion interactions may help elucidate the mechanism of how R. ruber cells obtain titanium from the metal-oxide particles.
Temple University--Theses
Williams, Andrea. "Characterisation of novel autophagy pathways : implications in the clearance of disease-causing aggregate-prone proteins." Thesis, University of Cambridge, 2008. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.612132.
Повний текст джерелаVincenz-Donnelly, Lisa Verfasser], та Franz-Ulrich [Akademischer Betreuer] [Hartl. "How the mammalian endoplasmic reticulum handles aggregation-prone β-sheet proteins / Lisa Vincenz-Donnelly ; Betreuer: Franz-Ulrich Hartl". München : Universitätsbibliothek der Ludwig-Maximilians-Universität, 2016. http://d-nb.info/1122019467/34.
Повний текст джерелаBorn, Ariane. "Etablierung und Optimierung der Error-Prone-PCR und eines Aktivitätsscreenings für Styrol-Monooxygenasen." Master's thesis, Technische Universitaet Bergakademie Freiberg Universitaetsbibliothek "Georgius Agricola", 2011. http://nbn-resolving.de/urn:nbn:de:bsz:105-qucosa-77143.
Повний текст джерелаStyrene monooxygenases (SMOs) play an important role in the bacterial degradation of styrene. They epoxidize the hydrocarbon highly enantioselective to (S)-styrene oxide. Most of the styrene monooxygenases known so far were identified in Gram-negative microorganisms like pseudomonads. Rhodococcus opacus 1CP, a Gram-positive nocardioform actinobacterium, which uses styrene as energy and carbon source was recently found to possess a novel type of SMO, StyA2B. This protein represents a natural fusion between an FAD:NADH oxidoreductase (StyB) and a single monooxygenase subunit (StyA2) and might act in combination with another single oxygenase StyA1 in strain 1CP. Two artificial analogs to StyA2B, designated StyAL1B and StyAL2B, were recently prepared by a fusion of styA and styB of Pseudomonas fluorescens ST and both showed oxygenating activity. For StyA1/StyA2B as well as the artificial fusion proteins StyAL1B and StyAL2B, it was tried to enhance the specific oxygenation activity in order to support their biotechnological applicability. The method of error prone PCR was used for that purpose. In order to identify favorable modifications with increased catalytic activity from a high number of mutants, an easy and simple screening test is necessary. Therefore, it is reasonable to use the ability of SMOs to oxidize indole to the blue dye indigo. However, the expression of SMOs as soluble proteins is an important requirement for any activity screening. Attempts to modify the genes styA2B and styA1/styA2B by means of a commercial error prone PCR kit yielded 300 to 1,200 potential mutants. Unfortunately, none of the obtained colonies showed any indole-oxidizing activity and the formation of insoluble inclusion bodies was assumed to be a likely explanation. In contrast to StyA2B and StyA1, recombinant expression of the artificial fused SMOs StyAL1B und StyAL2B should yield detectable amounts of active proteins. In fact, cultivation of clones expressing both types of proteins showed a blue coloration. Since the coloration of clones from one single solid medium evolved in a non-uniform manner, cultivation conditions were varied in order to identify factors which promote a more uniform tendency for indole oxidation. Although a high NaCl concentration in the medium was shown to favor indole oxidation, the latter one seems to be influenced by additional physiological factors, hardly to control. For the artificially fused proteins an error prone PCR was carried out, too. Although the initial step of mutagenic PCR was found to be successful, completing the vector system by a second ll-up PCR reaction failed. Alternative strategies like the usage of alternative DNA polymerases as well as a conventional cloning approach of various genes into a digested expression vector did not lead to detectable clones. The cultivation of identical clones on petri dishes provided no uniform tendency for indole oxidation and thus did not allow the reliable comparison of mutants in respect of their specific SMO activities. Cultivation of mutants in liquid medium should lead to more reproducible conditions and for that purpose a method was successfully established to quantify indigo formation and cell density
Liu, Xueyun, and Xueyun Liu. "Probe Ca2+/Camodulin reguation of membrane proteins engineering." Digital Archive @ GSU, 2013. http://digitalarchive.gsu.edu/chemistry_theses/59.
Повний текст джерелаMossuto, 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.
Повний текст джерелаSOUZA, Renata Silva Cabral de. "Avalia??o do potencial antioxidante e antimicrobiano de prote?nas do soro de leite concentradas por membranas e hidrolisadas por diferentes enzimas comerciais." Universidade Federal Rural do Rio de Janeiro, 2013. https://tede.ufrrj.br/jspui/handle/jspui/2534.
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CAPES
The aim of this study was to evaluate the process of protein concentration in bovine whey proteins by ultrafiltration process and subsequently the protein hydrolysate obtained by enzymatic hydrolysis to produce bioactive peptides with potential antimicrobial and antioxidant activities. For concentration process was used a ceramic ultrafiltration membrane with a molecular range cut-off of 10-20 kDa, transmembrane pressure of 5 bar and, temperature of 30 ?C, 40 ?C and 50 ?C . The optimum temperature condition was at 40 ?C. The Volume Concentrate Factor (VCF) parameter was used as a end-point of the ultrafiltration process and fixed at 2, corresponding on concentrating the initial volume twice, in volume. At the temperature of 40 ?C, VCF had a correspondence on final protein concentration on the concentrated fraction by ultrafiltration and confirmed by Bradford method. Two commercial enzymes were tested Alcalase, Flavourzyme and an equivalent mixture of both 50:50 (w/w) in the hydrolysis reaction. The hydrolysis conditions were determined according to manufacturer instructions and confirmed by other studies: 60 ?C and pH 8 for Alcalase; 50 ?C and pH 7 for Flavourzyme; 50 ?C and pH 8 for enzyme mixture with enzyme / substrate ratio (w / w) 5/100 for all enzymes. The reaction was monitored by pH Stat method. The final Degree of Hydrolysis (DH) achieved was 15%, 52% and 63% for Flavourzyme, Alcalase and enzyme mixture, respectively. Five aliquots were collected along the hydrolysis for each enzyme reaction corresponding to differents DH in order to evaluatethe antioxidant activity by ORAC and ABTS assays with final values between 597- 1092 m? TE (ABTS) and from 1615 to 2920 ?M TE (ORAC) for Flavourzyme; 998-6290 ?M TE (ABTS) and 3092-7567 ?M TE ( ORAC) for Alcalase and finally 913-2678 ?M TE (ABTS) and 2547-5903 ?M TE (ORAC) for the enzyme mixture. The samples from all hydrolysates showed no antimicrobial activity against strains of Salmonella choleraesuis subsp. Enteritidis (ATCC 13076) and Listeria monocytogenes (ATCC 9117).
A proposta do presente trabalho foi avaliar a concentra??o das prote?nas do soro de leite bovino por ultrafiltra??o e posterior obten??o de hidrolisados proteicos deste concentrado via hidr?lise enzim?tica visando obter pept?deos bioativos com potencial atividade antimicrobiana e antioxidante. Para concentra??o das prote?nas do soro foi utilizada membrana cer?mica de ultrafiltra??o com massa molar de corte de 10-20 kDa, press?o aplicada ? membrana de 5 bar, temperaturas testadas (30 ?C, 40 ?C e 50 ?C) . A temperatura ?tima selecionada foi de 40 ?C. O Fator de Concentra??o Volum?trica foi o par?metro utilizado para indicar o final do processo de ultrafiltra??o sendo fixado em duas vezes o volume inicial da alimenta??o. Na temperatura de 40 ?C foi obtida correspond?ncia entre a concentra??o volum?trica e a concentra??o proteica final na fra??o retida pela UF, que tamb?m foi o dobro da encontrada na fra??o alimenta??o, avaliada pelo m?todo de Bradford. Foram testadas duas enzimas comerciais: Alcalase, Flavourzyme e uma mistura equivalente de ambas, na propor??o 50:50 (m/m) na rea??o de hidr?lise. As condi??es de rea??o enzim?tica foram determinadas de acordo com instru??es do fabricante e corroboradas por outros estudos em: 60 ?C, pH 8 para Alcalase; 50 ?C, pH 7 para Flavourzyme; 50 ?C, pH 8 para mistura enzim?tica e rela??o enzima/substrato (g/g) foi de 5/100 para todas as enzimas. A rea??o de hidr?lise foi monitorada pelo m?todo pH Stat. Os Graus de Hidr?lise (GH) finais alcan?ados foram de 15 %, 52 % e 63 % para Flavourzyme, mistura enzim?tica e Alcalase, respectivamente. Foram coletadas cinco al?quotas correspondentes a diferentes GH ao longo da rea??o para cada condi??o enzim?tica utilizada e avaliadas quanto a atividade antioxidante pelos m?todos ABTS e ORAC tendo valores entre 597 a 1092 ?M TE (ABTS) e 1615 a 2920 ?M TE (ORAC) para Flavourzyme, 998 a 6290 ?M TE (ABTS) e 3092 a 7567 ?M TE (ORAC) para Alcalase e por fim, 913 a 2678 ?M TE (ABTS) e 2547 a 5903 ?M TE (ORAC) para a mistura enzim?tica. Nenhuma das amostras de hidrolisado com diferentes GH apresentou atividade antimicrobiana contra cepas de Salmonella choleraesuis subsp. Enteritidis (ATCC 13076) e Listeria monocytogenes (ATCC 9117).
Wikström, Malin. "Synthesis and protein curing abilities of membrane glycolipids." Doctoral thesis, Stockholm University, Department of Biochemistry and Biophysics, 2006. http://urn.kb.se/resolve?urn=urn:nbn:se:su:diva-1361.
Повний текст джерелаThere are many types of membrane lipids throughout Nature. Still little is known about synthesizing pathways and how different lipids affect the embedded membrane proteins. The most common lipids are glycolipids since they dominate plant green tissue. Glycolipids also exist in mammal cells as well as in most Gram-positive bacteria. Glycosyltransferases (GTs) catalyze the final enzymatic steps for these glycolipids. In the bacteria Acholeplasma laidlawii and Streptococcus pneumonie and in the plant Arabidopsis thaliana, GTs for mono-/di-glycosyl-diacylglycerol (-DAG) are suggested to be regulated to keep a certain membrane curvature close to a bilayer/nonbilayer phase transition. The monoglycosylDAGs are nonbilayer-prone with small headgroups, hence by themselves they will not form bilayer structures.
Here we have determined the genes encoding the main glycolipids of A. laidlawii and S. pneumonie. We have also shown that these GTs belong to a large enzyme group widely spread in Nature, and that all four enzymes are differently regulated by membrane lipids. The importance of different lipid properties were traced in a lipid mutant of Escherichia coli lacking the major (75 %), nonbilayer-prone/zwitterionic, lipid phosphatidylethanolamine. Introducing the genes for the GTs of A. laidlawii and two analogous genes from A. thaliana yielded new strains containing 50 percent of glyco-DAG lipids. The monoglyco-DAG strains contain significant amounts of nonbilayer-prone lipids while the diglyco-DAG strains contain no such lipids. Comparing these new strains for viability and the state of membrane-associated functions made it possible to connect different functions to certain lipid properties. In summary, a low surface charge density of anionic lipids is important in E.coli membranes, but this fails to be supportive if the diluting species have a too large headgroup. This indicates that a certain magnitude of the curvature stress is crucial for the membrane bilayer in vivo.
Robertson, Neil. "Development and application of simple FRET-based methods for aggregation-prone LIM domain interactions." Thesis, The University of Sydney, 2017. http://hdl.handle.net/2123/16912.
Повний текст джерелаTaylor, Christopher George. "Novel fluorescence techniques to probe protein aggregation." Thesis, University of Cambridge, 2018. https://www.repository.cam.ac.uk/handle/1810/276197.
Повний текст джерелаRobinson, Matthew D. "A novel fluorinated probe for medical imaging." Thesis, University of Oxford, 2014. http://ora.ox.ac.uk/objects/uuid:3f9e6bbf-bbda-45c3-9ff9-826463ff011e.
Повний текст джерелаSelvan, Nithya. "Reductionist animal models to probe protein O-GlcNAcylation." Thesis, University of Dundee, 2015. https://discovery.dundee.ac.uk/en/studentTheses/6034adb4-fc6a-4c47-ade6-74e0b18d4297.
Повний текст джерелаHabtemariam, Mesay. "Bioinformatics Approach to Probe Protein-Protein Interactions: Understanding the Role of Interfacial Solvent in the Binding Sites of Protein-Protein Complexes;Network Based Predictions and Analysis of Human Proteins that Play Critical Roles in HIV Pathogenesis." VCU Scholars Compass, 2013. http://scholarscompass.vcu.edu/etd/2997.
Повний текст джерелаCUNATI, DIANA. "Ruolo dei lipid raft nel metabolismo della proteina prionica." Doctoral thesis, Università degli Studi di Milano-Bicocca, 2011. http://hdl.handle.net/10281/27001.
Повний текст джерелаD'Angelo, Anna. "Functional characterization of the human PRUNE protein : implications in cancer." Thesis, Open University, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.406475.
Повний текст джерелаShadmehr, Mehrdad, and Mehrdad Shadmehr. "Design and Synthesis of Triazabutadiene-based Fluorogenic Probes for Tyrosine Specific Labeling of Proteins." Diss., The University of Arizona, 2018. http://hdl.handle.net/10150/626757.
Повний текст джерелаDehon, Patricia M. "Localization and Mutational Analysis of the Nuclear and Aggregation-Prone Ime4 Protein in Saccharomyces cerevisiae." ScholarWorks@UNO, 2012. http://scholarworks.uno.edu/td/1567.
Повний текст джерелаEgleton, James Edward. "Small molecule colorimetric and fluorescent probes for specific protein detection." Thesis, University of Oxford, 2015. http://ora.ox.ac.uk/objects/uuid:0a1a1c80-8055-491a-920a-3e17f7919e93.
Повний текст джерелаCaufield, J. Harry. "Interactomics-Based Functional Analysis: Using Interaction Conservation To Probe Bacterial Protein Functions." VCU Scholars Compass, 2016. http://scholarscompass.vcu.edu/etd/4580.
Повний текст джерелаThompson, Cherrie Delos Santos. "Using the potent Pasteurella multocida toxin to probe G-protein signalling." Thesis, King's College London (University of London), 2016. https://kclpure.kcl.ac.uk/portal/en/theses/using-the-potent-pasteurella-multocida-toxin-to-probe-gprotein-signalling(b9692597-dd37-4fbc-ba0d-00691809538e).html.
Повний текст джерелаQuintyn, Royston Samuel. "Applying Tandem Mass Spectrometry coupled with Ion Mobility to probe the Structure of Non-Covalent Protein Complexes and their Interactions with Ligands, Peptides and other Proteins." The Ohio State University, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=osu1437495530.
Повний текст джерелаSilva, Kássya Mycaela Paulino da. "Avaliação do Nível Basal e do Controle Barorreflexo da Atividade do Sistema Nervoso Simpático da Prole de Ratos que Sofreram Desnutrição Proteica Durante a Gestação e a Lactação." Universidade Federal de Pernambuco, 2015. https://repositorio.ufpe.br/handle/123456789/16439.
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FACEPE
Estudos buscam esclarecer a relação entre desnutrição proteica em fase crítica do desenvolvimento e surgimento de hipertensão arterial na vida adulta, entretanto os mecanismos que participam deste processo ainda não são totalmente conhecidos. Neste estudo investigamos as repercussões da desnutrição proteica perinatal (gestação e lactação) sobre o nível basal e o controle barorreflexo da atividade do sistema nervoso simpático da prole. Ratas prenhes foram alimentadas com dieta hipoproteica (8% de proteína, grupo experimental) ou com dieta normoproteica (17% de proteína, grupo controle) durante o período perinatal. Após o desmame, a prole de ratos machos recebeu dieta padrão para animais de laboratório e os estudos funcionais foram realizados aos 90 dias de vida. Todos os protocolos experimentais foram aprovados pelo comitê de ética em utilização animal da UFPE (processo nº 23076.016256/2013-83). Para registro direto das variáveis hemodinâmicas e infusão das drogas, foram implantadas cânulas na artéria e veia femoral, respectivamente. Os níveis de atividade autonômica foram estimados através de métodos lineares e não lineares de análise da variabilidade da pressão sistólica e intervalo de pulso em animais conscientes e anestesiados, enquanto a eficiência do mecanismo barorreflexo em regular a pressão arterial foi avaliado através de métodos indiretos de sequência espontânea e de forma induzida mediante infusão de fenilefrina e nitroprussiato de sódio. Além disso, foi realizado implante de eletrodo bipolar na cadeia simpática em nível L3-L5 para registro direto da atividade elétrica do nervo lombar em animais anestesiados. Os resultados demonstram que ratos submetidos à desnutrição proteica perinatal apresentam desbalanço autonômico para o coração, com aumento do componente de baixa frequência no espectro da pressão sistólica e intervalo de pulso, aumento do índice simpatovagal para o coração e aumento dos padrões 0V, relacionados ao tônus simpático cardiovascular. Além disso, observamos a integridade do controle barrorreflexo da pressão arterial em todas as doses avaliadas, uma vez que os animais não anestesiados submetidos à restrição proteica apresentaram sensibilidade barorreflexa semelhante aos controles. Em condições de anestesia, o registro direto do nervo lombar em ratos desnutridos não foi diferente dos seus pares, demonstrando que o controle barorreflexo da atividade nervosa simpática lombar está preservado neste modelo experimental. Esses achados apontam que o desenvolvimento da hipertensão arterial na prole submetida à desnutrição proteica perinatal não está relacionada à disfunção do barorreflexo em animais conscientes e anestesiados. Sugerimos que outros mecanismos centrais e periféricos possam estar envolvidos no surgimento e manutenção dos níveis elevados de pressão arterial neste modelo experimental.
Studies seek to clarify the relationship between protein malnutrition in critical stage of development and emergence of hypertension in adulthood, however the mechanisms involved in this process are not yet fully known. We investigated the effects of perinatal protein malnutrition (pregnancy and lactation) on the baseline and the baroreflex control of the sympathetic nervous system activity on the offspring. Pregnant rats were fed a low protein diet (8% protein, experimental group) or normal protein diet (17% protein, control group) during the perinatal period. After weaning, the male offspring of rats received standard diet to laboratory animals and functional studies were performed at 90 days of life. All experimental protocols were approved by the Ethics Committee on Animal use of UFPE (process n° 23076.016256 / 2013-83). For the direct recording of hemodynamic variables and for the infusion of drugs, cannulas were implanted in the femoral artery and vein, respectively. The autonomic activity levels were estimated by linear and nonlinear methods of analysis of the variability of systolic and pulse interval in conscious and anesthetized animals. The efficiency of the baroreflex mechanism in regulating the blood pressure was evaluated by indirect methods of sequence and form spontaneously induced by phenylephrine and sodium nitroprusside infusion. We also performed bipolar electrode implantation in the sympathetic chain at L3-L5 level for direct recording of the electrical activity of the lumbar nerve in anesthetized animals. The results show that rats with perinatal protein malnutrition have autonomic imbalance to the heart, with increased low frequency component in the spectrum of systolic and pulse interval, increased sympathovagal index to the heart and increase the standards 0V, related to the tone cardiovascular friendly. In addition, we observe the integrity of baroreflex control of blood pressure in all evaluated doses, since unanesthetized animals submitted to protein restriction had baroreflex sensitivity similar to controls. In anesthesia conditions, the record of straight lumbar nerve in malnourished rats was not different from their peers, demonstrating that the baroreflex control of lumbar sympathetic nerve activity is preserved in this experimental model. These findings indicate that the development of hypertension in offspring submitted to perinatal protein malnutrition is not related to the baroreflex dysfunction in conscious and anesthetized animals. We suggest that other central and peripheral mechanisms may be involved in the emergence and maintenance of high levels of blood pressure in this experimental model.
Rodrigues, Mariane Augusta Domingues. "Avaliação da atividade e resistência à clivagem proteolítica de L-asparaginases recombinantes obtidas por reação em cadeia da polimerase propensa a erro." Universidade de São Paulo, 2016. http://www.teses.usp.br/teses/disponiveis/9/9134/tde-24082016-163433/.
Повний текст джерелаEscherichia coli L-asparaginase (EcA II) is an enzyme widely used in the treatment of acute lymphoblastic leukemia (ALL), acting in the depletion of the amino acid L-asparagine, which is essential for cancer cells proliferation. However, treatment with L-asparaginase is associated with a high rate of hypersensitivity, due to formation of anti-L-asparaginase antibody and the enzyme cleavage by the serum proteases asparagine endopeptidase (AEP) and cathepsin B (CTSB). Furthermore, the neurotoxicity is associated with the effect of the enzyme L-glutaminase activity. The main aim of the current work is to obtain variants of EcA II (gene ansB) with an equivalent catalytic efficiency, greater resistance to proteolytic cleavage and a reduced glutaminase activity. For such purpose, through error-prone polymerase chain reaction (epPCR) of gene ansB, a library of 1128 clones was constructed in pET15b vector and expressed in BL21(DE3). None mutant with an asparaginase activity equivalent to EcA II wild type showed a reduced glutaminase activity. Among the screened clones, one mutant (T161I) was resistant to CTSB proteolytic cleavage and two mutants (Q190L e P40S/S206C) were resistant to both CTSB and AEP proteolytic cleavages. These three mutants were EcA II wild type equivalents in asparaginase and glutaminase activities. Our data show promising new possibilities of mutant EcA II presenting higher stability against human serum proteolytic cleavage and maybe lower immunogenicity.
Laos, Roberto. "Protein directed evolution." Revista de Química, 2012. http://repositorio.pucp.edu.pe/index/handle/123456789/99875.
Повний текст джерелаDirected evolution allows us to explore protein functionalities not required in the natural environment. It mimics natural genetic processes and selective pressures. This approach is used when the molecular basis is not completely understood and rational design is a difficult task. This approach consists of serial cycles of consecutive diversification and selection which eventually lead to the accumulation of beneficial mutations. Here are presented two cases where directed evolution is used to modify two different proteins: Taq polymerase, enzyme used for DNA extension in PCR, and the LacI repressor protein which regulates gene expression on E.coli.
Clark, Peter George Keith. "Discovery of epigenetic probes against the bromodomain family of proteins." Thesis, University of Oxford, 2015. https://ora.ox.ac.uk/objects/uuid:a589d1f3-c8f6-4219-a10b-79099eafe1c8.
Повний текст джерелаStofella, Michele. "Hydrogen deuterium exchange: methods to probe protein dynamics at single residue resolution." Master's thesis, Alma Mater Studiorum - Università di Bologna, 2020. http://amslaurea.unibo.it/21242/.
Повний текст джерелаHuisstede, Jurgen Hendrikus Gerhardus. "Scanning probe optical tweezers a new tool to study DNA-protein interactions /." Enschede : University of Twente [Host], 2006. http://doc.utwente.nl/55834.
Повний текст джерелаChen, Changsheng Verfasser], and Maximilian [Akademischer Betreuer] [Ulbrich. "Fluorescently labeled DNA probe in STORM imaging and single-molecule protein labeling." Freiburg : Universität, 2019. http://d-nb.info/1187658545/34.
Повний текст джерелаChen, Changsheng [Verfasser], and Maximilian [Akademischer Betreuer] Ulbrich. "Fluorescently labeled DNA probe in STORM imaging and single-molecule protein labeling." Freiburg : Universität, 2019. http://d-nb.info/1187658545/34.
Повний текст джерелаKhan, Md Ahad Ali. "Molecular weight information from protein-ligand complexes to probe natural product interactions." Thesis, Griffith University, 2017. http://hdl.handle.net/10072/371908.
Повний текст джерелаThesis (PhD Doctorate)
Doctor of Philosophy (PhD)
School of Natural Sciences
Science, Environment, Engineering and Technology
Full Text
Mosconi, Francesco. "Fluctuations in biological molecules : tools to probe mechanical and structural properties of DNA and proteins." Paris 6, 2008. http://www.theses.fr/2008PA066344.
Повний текст джерелаMosconi, Francesco. "Fluctuations in biological molecules: tools to probe mechanical and structural properties of DNA and proteins." Doctoral thesis, Università degli studi di Padova, 2008. http://hdl.handle.net/11577/3426746.
Повний текст джерелаCanale, Vinicius 1986. "Estudo funcional e morfológico renal da prole de ratos cujas mães foram submetidas à restrição proteica gestacional : efeito do tratamento com rapamicina." [s.n.], 2014. http://repositorio.unicamp.br/jspui/handle/REPOSIP/312741.
Повний текст джерелаDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas
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Resumo: Já está bem documentado que a dieta hipoproteica em ratos durante a gestação produz efeitos no crescimento fetal, uma persistente elevação na pressão arterial e disfunções no desenvolvimento renal da prole. Alterações na pressão arterial parecem estar relacionadas à acentuada redução no número de nefros que acaba por causar um quadro de hipertrofia e hiperfluxo nos nefros remanescentes como adaptação para equilibrar a taxa de filtração, no entanto, os mecanismos utilizados para esta adaptação culminam com o surgimento de albuminúria. Este processo acaba por causar esclerose glomerular culminando em um ciclo, comprometendo cada vez mais os nefros remanescentes. O desenvolvimento deste quadro pode levar à síndrome nefrótica e posteriormente à doença renal terminal. Acredita-se que o principal fator atuante na programação fetal neste modelo é devido à exposição exacerbada do feto aos glicocorticoides materno, o que acaba por comprometer a correto desenvolvimento e diferenciação de tecidos e órgãos, e na expressão ou atividade de uma série de receptores e enzimas. Recentemente, tem surgido a hipótese de que a atividade da mTORC poderia estar envolvida no surgimento de doenças na idade adulta neste modelo experimental. Este trabalho teve por objetivo avaliar se a inibição da mTORC através do tratamento com rapamicina em animais programados poderia ser benéfico, inibindo o surgimento de complicações relacionadas com a estrutura glomerular e como seria seu efeitos sistêmico, sobre a função renal e pressão arterial sistêmica. Ratos Wistar receberam ração com baixa proteína (6% LP) e dieta controle (17% NP) durante o período gestacional. A prole de machos foi tratada com rapamicina diluída em DMSO (5%) e administrada via intraperitoneal na dose de 1mg/kg, 3 vezes por semana, a partir da 4ª semana de vida até a 12ª semana. A aferição da pressão arterial sistólica foi realizada nas idades 8, 12 e 16. Foi observado que nos grupos que receberam rapamicina, a pressão artéria sistólica elevou-se consideravelmente em todas as idades. A avaliação da função renal foi realizada através de clearance de creatinina e lítio nas mesmas idades e observamos que durante todo o tratamento, o grupo NP que recebeu rapamicina excretou mais sódio, na porção pós-proximal do túbulo. Além disso, não houve diferença na taxa da filtração glomerular em nenhuma das idades. Quando a proteinúria foi avaliada, observamos que o grupo programado LP sem rapamicina, apresentou evolução com o passar das semanas sendo significativamente maior a partir das 12ª e 16ª semanas, no entanto, os grupos que receberam rapamicina, não apresentaram a mesma evolução, indicando preservação da estrutura glomerular. Os presentes resultados demonstram que apesar de a rapamicina ter elevado a pressão arterial em ambos os grupos, há uma indicação de que os animais programados tem um controle menos eficaz no controle da pressão arterial através da função renal. Mesmo diante da pressão elevada, a rapamicina foi capaz de inibir injúrias à barreira de filtração
Abstract: It is well established that a low protein diet in rats during pregnancy causes effects on fetal growth, a persistent elevation in blood pressure and renal dysfunction in the offspring at a later age. Alterations in blood pressure seem to be related to the marked reduction in the number of nephrons which ultimately causes an overflow and hypertrophy in the remaining nephrons in the attempt to adapt its balance on the filtration rate. However, the mechanisms used for this adaptation results in the appearance of albuminuria. This process might cause glomerular sclerosis culminating in a cycle, increasing the damage to the remaining nephrons. The development of this framework can lead to nephrotic syndrome and subsequently to ESRD. It is believed that the main factor in the fetal programming model is the fetus overexposure to maternal glucocorticoids, which ultimately compromises the proper development and differentiation of tissues and organs. Additionally, the expression and the activity of a number of receptors and enzymes is also affected. Recently, in this experimental model, there has arisen the hypothesis that the activity of mTORC could be involved in the onset of disease in adulthood. This study assess whether the inhibition of mTORC, through the treatment with rapamycin in programmed animals, could be beneficial by inhibiting the onset complications related to glomerular structure and its affect on renal function and blood pressure. Wistar female rats were fed with low protein (6 % LP) or control diet (17 % NP) during pregnancy. The male offspring were treated with rapamycin diluted in DMSO (5 %) and administered intraperitoneally at a dose of 1mg/kg, 3 times per week, from the 4th week of life until the 12th week. The measurement of systolic blood pressure was measured at 8, 12, and 16 weeks old. We noticed that in the groups treated with rapamycin the arterial systolic pressure rose considerably in all ages. The assessment of renal function was performed by creatinine clearance and lithium at the same ages as the blood pressures assessment. The NPR group that received rapamycin had an increase in sodium excretion at the post- proximal tubule during the whole treatment. Additionally, there was no difference in the rate of glomerular filtration rate at any age among the groups. When proteinuria was assessed, we found that the programmed Group LP without rapamycin, showed an increase along the weeks and as expected, the groups that have received rapamycin did not show the same trend, indicating preservation in the glomerular structure. The present results demonstrate that rapamycin caused increase in the blood pressure in both groups however, it was able still to inhibit the injury into the filtration barrier. Additionally, there is an indication that the programmed animals, has a less effective control in the blood pressure and the excretion of sodium, even when the treatment is interrupted
Mestrado
Fisiopatologia Médica
Mestre em Ciências
McDonagh, Paul David. "Protein engineering to probe the catalytic mechanism of Alpha-class glutathione S-transferases." Thesis, University of Leicester, 1997. http://hdl.handle.net/2381/29679.
Повний текст джерелаSun, Li. "Advances in the use of phosphorescence spectroscopy as a probe of protein flexibility." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1997. http://www.collectionscanada.ca/obj/s4/f2/dsk3/ftp04/nq30395.pdf.
Повний текст джерелаLi, Xin. "Development and Application of Chemical and Structural Biology Approaches to Probe Protein Function." The Ohio State University, 2011. http://rave.ohiolink.edu/etdc/view?acc_num=osu1306439016.
Повний текст джерелаWorthman, Lynn-Ann D. "Surfactant protein A (SP-A) affects pulmonary surfactant morphology and biophysical properties." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1997. http://www.collectionscanada.ca/obj/s4/f2/dsk2/tape16/PQDD_0014/MQ34241.pdf.
Повний текст джерелаSagle, Laura B. "Carbon-deuterium bonds as an infrared probe of protein dynamics, local electrostatics and folding." Connect to a 24 p. preview or request complete full text in PDF format. Access restricted to UC campuses, 2006. http://wwwlib.umi.com/cr/ucsd/fullcit?p3195630.
Повний текст джерелаTitle from first page of PDF file (viewed February 28, 2006). Available via ProQuest Digital Dissertations. Vita. Includes bibliographical references.
Shearer, Lee. "Design and synthesis of ligands to probe the interactions of retinol binding protein (RBP)." Thesis, University of Leeds, 2011. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.713694.
Повний текст джерелаSowley, Hugh Richard. "Electron-vibration-vibration two-dimensional infrared spectroscopy as a structural probe of interactions in proteins and DNA." Thesis, Imperial College London, 2017. http://hdl.handle.net/10044/1/57959.
Повний текст джерелаGantner, Matthias. "Exploring solid supported membrane based electrophysiology as an alternative platform to probe activity of membrane transport proteins." Thesis, University of Leeds, 2017. http://etheses.whiterose.ac.uk/19682/.
Повний текст джерелаNacheva, Katya Pavlova. "Development of a Bio-Molecular Fluorescent Probe Used in Kinetic Target-Guided Synthesis for the Identification of Inhibitors of Enzymatic and Protein-Protein Interaction Targets." Scholar Commons, 2012. http://scholarcommons.usf.edu/etd/4376.
Повний текст джерелаBychkova, A. V., O. N. Sorokina, P. G. Pronkin, A. S. Tatikolov, A. L. Kovarski, and M. A. Rosenfeld. "Protein-Coated Magnetic Nanoparticles: Creation and Investigation." Thesis, Sumy State University, 2013. http://essuir.sumdu.edu.ua/handle/123456789/35378.
Повний текст джерелаSorin, Eric J. "Biomolecular assembly in silico using massively parallel simulations to probe protein and RNA folding dynamics /." May be available electronically:, 2007. http://proquest.umi.com/login?COPT=REJTPTU1MTUmSU5UPTAmVkVSPTI=&clientId=12498.
Повний текст джерелаCustódio, Augusto Henrique 1983. "Estudo da denervação renal bilateral e da imunorreatividade para substância P (SP), CGRP e receptor 1 para neurocinina (NK1R) no gânglio da raiz dorsal e parede pélvica renal na prole de ratas submetidas a restrição proteica gestacional." [s.n.], 2014. http://repositorio.unicamp.br/jspui/handle/REPOSIP/312740.
Повний текст джерелаDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas
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Resumo: A programação fetal é um processo fisiológico que assegura, durante o desenvolvimento intrauterino, a adaptação para o mundo exterior. Ou seja, o organismo é "moldável" por estímulos durante sua formação e dependendo do insulto experimentado, existe a possibilidade de mudanças estruturais e funcionais que podem predispor o indivíduo a doenças na vida adulta. O modelo de restrição proteica, assim como outros modelos, leva a um "stress" gestacional e, segundo a hipótese de Barker, programa a prole ao desenvolvimento de doenças na vida adulta, dentre elas a hipertensão arterial. Os rins são órgãos fundamentais na manutenção do equilíbrio hemodinâmico. Mudanças morfológicas e neuroendócrinas nos rins levam a alterações hidroeletrolíticas frequentemente associadas à patogênese da hipertensão arterial. A gênese desta doença ainda não está bem descrita, por envolver alterações multifatoriais, dentre elas, modificações da atividade neural tanto central quanto periférica, que podem ser um indicativo da elevação pressórica em nosso modelo. A atividade simpática renal é um importante modulador da excreção dos eletrólitos e, quando alterada, promove maior ou menor retenção de sais, principalmente o sódio, podendo contribuir para a elevação da volemia e consequentemente da hipertensão arterial. Diversos neuropeptídeos estão envolvidos na atividade simpática renal e os níveis destes são um importante marcador na gênese da hipertensão. Dentre esses peptídeos estão a Substância P (SP), seu receptor NK1R e o Peptídeo Relacionado ao Gene da Calcitonina (CGRP). Nossos resultados mostraram redução na imunorreatividade de SP, CGRP e aumento do receptor 1 para neurocinina nos gânglios da raiz dorsal da prole de ratas submetidas à restrição proteica gestacional. Identificamos também a elevação dos níveis de CGRP na parede pélvica renal. Assim, acreditamos que haja alterações na neuromodulação da atividade aferente renal, o que pode ser um fator contribuinte para a manutenção do estado hipertensivo neste modelo experimental
Abstract: A fetal programming is a physiologic process that ensures an adaptation for external world during the intra uterine development. In this period, the organism is "moldable" by stimulus that happens during its formation, which ensures adequate phenotypes formation for different environments. Kidneys are the most important organs when it has to do with maintaining the organism hemodynamic balance and also morphological and neuroendocrine alterations, which leads to fluid and eletrolytes changes, frequently associated to arterial hypertension pathogenesis. The genesis of this disease is not well described yet. It involves multifactorial changes like the neural activity in both central as peripheral, which, in our model, may be an indicative of increased pressure. The renal sympathetic activity is an important excretion modulator of electrolytes and when amended, promotes greater or lesser retention of salts, mainly sodium, contributing to the increase in blood volume and consequently hypertension. Several neuropeptides are involved in renal sympathetic activity, and these levels are an important marker in the genesis of hypertension. Among these peptides we find substance P (SP) and its receptor NK1R, and Related Peptide Calcitonin Gene (CGRP). Our research showed reduced immunoreactivity of SP, CGRP and increased neurokinin 1 receptor in dorsal root ganglia among the offspring of rats subjected to gestational protein restriction. According to this result, we believe that there are changes in afferent renal activity neuromodulation which may be a contributing factor for maintenance of hypertension in this experimental model
Mestrado
Medicina Experimental
Mestre em Ciências
Hanyu, Yuki. "Chemical scanning probe lithography and molecular construction." Thesis, University of Oxford, 2010. http://ora.ox.ac.uk/objects/uuid:409308ed-4806-44fc-87c3-5c1fe8971f79.
Повний текст джерелаLee, Peter S. M. Massachusetts Institute of Technology. "Using optical tweezers, single molecule fluorescence and the ZIF268 protein-DNA system to probe mechanotransduction mechanisms." Thesis, Massachusetts Institute of Technology, 2005. http://hdl.handle.net/1721.1/34490.
Повний текст джерелаIncludes bibliographical references (p. 42-43).
Optical tweezers instruments use laser radiation pressure to trap microscopic dielectric beads. With the appropriate chemistry, such a bead can be attached to a single molecule as a handle, permitting the application of force on the single molecule. Measuring the force applied in real-time is dependent on detecting the bead's displacement from the trapping laser beam axis. Back-focal-plane detection provides a way of measuring the displacement, in two-dimensions, at nanometer or better resolution. The first part of this work will describe the design of a simple and inexpensive position sensing module customized for optical tweezers applications. Single molecule fluorescence is another powerful technique used to obtain microscopic details in biological systems. This technique can detect the arrival of a single molecule into a small volume of space or detect the conformational changes of a single molecule. Combining optical tweezers with single-molecule fluorescence so that one can apply forces on a single molecule while monitoring its effects via single molecule fluorescence provides an even more powerful experimental platform to perform such microscopic studies. Due to the enhanced photobleaching of fluorophores caused by the trapping laser, this combined technology has only been demonstrated under optimized conditions.
(cont.) The second part of this work will describe a straightforward and noninvasive method of eliminating this problem. The study of mechanotransduction in biological systems is critical to understanding the coupling between mechanical forces and biochemical reactions. Due to the recent advances in single molecule technology, it is now possible to probe such mechanisms at the single molecule level. The third and final part of this work will describe a basic mechanotransduction experiment using the well-studied ZIF268 protein-DNA system. An experimental assay and method of analysis will be outlined.
by Peter Lee.
S.M.
Neely, Robert K. "The photophysical properties of 2-aminopurine and its application as a probe of DNA-protein interactions." Thesis, University of Edinburgh, 2005. http://hdl.handle.net/1842/12712.
Повний текст джерела