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Статті в журналах з теми "Caratterizzazione strutturale e biochimica"
Barreca, L., PA Marziliano, G. Menguzzato, and A. Scuderi. "Stand structure and dead wood characterization in cork forest of Calabria region (southern Italy)." Forest@ - Rivista di Selvicoltura ed Ecologia Forestale 7, no. 1 (July 30, 2010): 158–68. http://dx.doi.org/10.3832/efor0628-007.
Повний текст джерелаDi Berardino, Claudio, and Giuseppe Mauro. "Crescita economica e impatto della crisi: il ruolo dei distretti industriali in Italia." ECONOMIA E SOCIETÀ REGIONALE, no. 1 (June 2011): 92–114. http://dx.doi.org/10.3280/es2011-001009.
Повний текст джерелаOlivito, Renato S., Saverio Porzio, Carmelo Scuro, Domenico Luca Carnì, and Francesco Lamonaca. "Inventory and monitoring of historical cultural heritage buildings on a territorial scale: a preliminary study of structural health monitoring based on the CARTIS approach." ACTA IMEKO 10, no. 1 (March 31, 2021): 57. http://dx.doi.org/10.21014/acta_imeko.v10i1.820.
Повний текст джерелаДисертації з теми "Caratterizzazione strutturale e biochimica"
Liberi, Stefano <1990>. "CARATTERIZZAZIONE STRUTTURALE DELL’ALBUMINA SIERICA UMANA IN COMPLESSO CON UN CONTAMINANTE AMBIENTALE." Master's Degree Thesis, Università Ca' Foscari Venezia, 2020. http://hdl.handle.net/10579/16297.
Повний текст джерелаMontefusco, Sandro. "Studio del ruolo funzionale e caratterizzazione strutturale del cuprocomplesso TFF1-Cu." Doctoral thesis, Universita degli studi di Salerno, 2012. http://hdl.handle.net/10556/1311.
Повний текст джерелаThe maintenance of gastrointestinal tissue integrity is physiologically essential in the presence of the persistent harassment of microbial flora and injurious agents. Even though the repair of the gastric epithelium may be modulated by several factors, the epithelial continuity also depends on a family of small peptides called trefoil factors (TFFs). The trefoil factors family comprises the gastric peptides pS2/TFF1, the spasmolytic peptide (SP)/TFF2 and the intestinal trefoil factor (ITF)/TFF3; they are characterized by a three looped domain, the “trefoil domain”, stabilized by three disulphide bridges. TFF1 and TFF3 also have a seventh cysteine that allows the formation of omo- and/or hetero-dimers. On the other hand TFF2 presents only a monomeric form, containing two trefoil domains in the same polypeptide chain. TFFs are small protease-resistant proteins that are abundantly produced by mucus-secreting cells of the gastrointestinal tract onto the mucosal surface. TFFs are essential in the protection of the mucosal epithelia against a wide range of biological threats, thus contributing to the mucosal repair. The signaling events that mediate the cellular responses elicited by TFFs are only partially understood. Moreover there are convincing evidence that TFFs do play an important role in tumorigenesis, even though their specific roles in cancer are still unclear. TFF1 expression is strongly induced after mucosal injury and it has been proposed that TFF1 functions as a gastric tumor suppressor gene. Several studies confirm that TFF1 expression is frequently lost in gastric cancer because of deletions, mutations or methylation of the TFF1 gene. Infection by Helicobacter pylori, a class 1 carcinogen according to WHO classification, is thought to promote stomach carcinogenesis through induction of aberrant DNA methylation. Samples from infected patients show lower expression of TFF1. Recent studies have also shown that there is a direct relationship between Helicobacter pylori and the dimeric form of the protein. In fact, it was demonstrated that the core oligosaccharide portion of H. pylori lipopolysaccharide (RF-LPS) is able to bind to TFF1. It also seems that the loss of TFF1 is an important event in shaping the NF-kB-mediated inflammatory response during the progression to gastric tumorigenesis, being TFF1 a negative regulator of NF-kB signalling. It is thus emerging a clear correlation between loss of TFF1, the development of inflammatory disease and the neoplastic process. Recent analyses made by our research group allowed us to point out the up-regulation of TFF1 gene expression in rats fed on copper deficient diets, and allowed us to find out the unexpected ability of TFF1 to bind copper ions. The presence of a cysteine surrounded by several negatively charged residues in the carboxy-terminus of the protein suggested the presence of a copper-binding site. Afterwards, it was shown that Cys58 and at least three Glu surrounding residues are essential to efficiently bind copper. Moreover, the incubation of the native peptide with copper salts increases the fraction of peptide omodimers produced by inter-molecular oxidation of Cys58 and disulphide bond formation. The Ph.D. research project was aimed at characterising the structure-function relationship of the TFF1-Cu complex. Briefly, we studied the influence of copper on known TFF1 biological activities and on its gene regulation, then we investigated its involvement in the TFF1 mediated mechanisms of Helicobacter pylori virulence and infection. A preliminary Real Time PCR quantitative analysis showed that copper deficiency positively modulates tff1 expression in an adenocarcinoma cell line (AGS), thus confirming our previous data obtained in vivo in copper deficient rat intestine. In order to map possible copper responsive elements in the proximal promoter sequence, we analysed the expression of a reporter gene (Luciferase) driven by deletion constructs of the tff1 gene promoter. AGS cells transfected with the deletion constructs allowed us to identify the upstream 5’ gene sequence -583/-435 as a promoter region sensitive to the changes of copper concentration. In fact, copper chelation treatments with bathocuproine disulfonate (BCS) were able to stimulate an increase of the promoter activity of the corresponding deletion construct. Following the sequence analysis (Transfac software) we focused our attention on a putative SP1 binding site identified in this region, whose binding ability was then confirmed by electrophoretic mobility shift assay (-561/-552). In agreemente with our in vitro results, it was also observed that copper favours the native TFF1 dimer formation in the culture medium of MCF-7 and HT29-MTX cells (a mucus-secreting clone obtained from the HT29 colon cancer cell line), thus confirming a possible role of the metal in the balance between the monomeric and the dimeric forms To evaluate the effect of copper-TFF1 interaction on the well known motogenic activity of the protein, we performed wound healing assays on an inducible clone of gastric cancer cells (AGS) able to overexpress the peptide (AGS-AC1). As expected, the overexpression of TFF1 stimulates an appreciable increase of cell migration, and copper chelation (BCS) undo the benefits of the increased peptide level. Our previous results showed that copper treatments decreased the amount of secreted protein in culture medium. Further experiments demonstrated that induced AGS-AC1 cells are able to store intracellularly higher amount of copper if compared to uninduced AGS-AC1 cells. This evidence suggests that TFF1 levels may also play a role also in the uptake/traffic of copper in this in vitro model. Finally, we studied the combined influence of TFF1 and copper in Helicobacter pylori infections. Our results demonstrate that Cu-TFF1 complex promotes H. pylori colonization of AGS cells. In fact, AGS-AC1 cells overexpressing TFF1 are more efficiently colonised by H. pylori wild-type (str. P12) if compared to uninduced cells. The presence of copper in a duplicate experiment further increases the colonization, as well as copper chelation by bathocuproine disulfonate (BCS) reduces the observed effect. The same result was obtained with H. pylori str. P12Δ479, an isogenic mutant expressing a truncated LPS core still able to bind to TFF1. On the other hand, H. pylori P12Δ1191, unable to bind TFF1, is not affected by copper levels in the culture medium. Parallel experiments were carried out on mucus secreting HT29-E12 goblet cells, to compare and/or confirm the results obtained in AGS-AC1. The results show that also in HT29-E12 cells the H. pylori colonization follows a similar trend, increasing when incubated in the presence of Cu and decreasing after BCS treatment. The present work contributed interesting results in the field of the biochemistry of the epithelia, in the wake of the research in progress in our laboratory aimed at studying the biological activities of the newly identified metalloprotein Cu-TFF1, whose properties are still poorly characterized. On the basis of the previous structural pieces of evidence we observed that the protein level and the balance of its oligomeric forms can be affected and regulated by copper ions. In turn, this delicate equilibrium is able to affect the integrity and the rheological properties of the epithelial barrier, thus representing a fine tuner, or an Achille’s heel, through which pathogenic microrganisms and deregulated proliferation of neoplastic cells may take advantage for their invasiveness. The role of copper in the TFFs biochemistry represents a new finding in the puzzling and versatile functions of this interesting peptide family, whose thorough comprehension still reserves many questions and surprises. [edited by author]
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Pucci, Kathleen. "Caratterizzazione strutturale e funzionale di proteine-effettori di patogenesi appartenenti alla "PcF Toxin Family"." Doctoral thesis, Università Politecnica delle Marche, 2009. http://hdl.handle.net/11566/242344.
Повний текст джерелаVETTRAINO, CHIARA. "O-fosfoetanolamina fosfo-liasi: dalla caratterizzazione strutturale ad applicazioni biomediche." Doctoral thesis, Università degli studi di Modena e Reggio Emilia, 2020. http://hdl.handle.net/11380/1200057.
Повний текст джерелаEnzymes have been used in many productive sectors for more than a hundred years. However, over the last 50 years, since the advent of recombinant DNA technologies, they have found a much wider and diverse range of applications, spanning across virtually any kind of human activities. In particular, because of their remarkable properties, enzymes have become the major choice for medical diagnostic applications and are being increasingly utilized for both the detection and the treatment of a broad variety of diseases. The purpose of this thesis is to study and exploit the high substrate specificity of a very peculiar human PLP-dependent enzyme, named O-phosphoethanolamine phospho-lyase (ETNPPL), for diagnostic applications. The enzyme promotes the irreversible degradation of O-phosphoethanolamine (PEA) to acetaldehyde, phosphate and ammonia. The physico-chemical characterization of ETNPPL reported in the literature suggests a mechanism of action that is very similar to other PLP-dependent enzymes, implying that the same residues are present in the active site. However, a structural characterization of human ETNPPL has not yet been reported in the literature. The first issue that I addressed in this thesis was to obtain the high-resolution 3D-structure of ETNPPL by X-ray crystallography in order to validate the mechanism of action proposed in the literature. To this end, starting from the cloning of the DNA fragment encoding for the human ETNPPL into a suitable expression vector, I expressed and purified the human ETNPPL. After assessing the enzymatic activity of the purified protein sample, I carried out crystallization experiments and I obtained good quality crystals for X-ray data collection. I determined the 3D-structure of ETNPPL at 2.3 Å resolution. Once I have completed the structural characterization of the enzyme and I have fully rationalized its high substrate specificity for phosphoethanolamine (PEA), its natural substrate, I set out to investigate one of its possible practical applications by designing and working towards the development of a biosensor for the selective detection of PEA in the urine. PEA levels in the urine of prostate cancer patients have been reported to decrease relative to healthy patients. Therefore, an ETNPPL-based assay for the measurement of PEA concentrations would be most useful in prostate cancer screening. To date, the early detection of prostate cancer involves the prostate-specific antigen (PSA) blood test. However, this approach is known to produce high rates of false positive results since increased PSA levels are often found in men with benign prostatic hyperplasia (BPH), prostatitis and prostate injury. For this reason, the need for new biomarkers that can be used for diagnostic purposes either alone or in combination with standard PSA tests appears evident. A method for the rapid and quantitative detection of this analyte would be of relevance for the diagnosis of such disease and, at the same time, would represent a potent validation tool of PEA as a prostate cancer biomarker.
GALASSI, LUCIA. "Regolazione del metabolismo del NAD nell'uomo: caratterizzazione cinetica, funzionale, regolatoria e strutturale della nicotinato fosforibosiltrsferasi (NaPRT)." Doctoral thesis, Università Politecnica delle Marche, 2011. http://hdl.handle.net/11566/241869.
Повний текст джерелаNicotinate phosphoribosyltransferase (NaPRT) catalyzes the conversion of nicotinate (Na) to Na mononucleotide (NaMN), the first reaction of the Preiss-Handler pathway for the biosynthesis of nicotinamide adenine dinucleotide (NAD). The importance of NaPRT in NAD metabolism is demonstrated by the fact that, in human embryonic kidney cells, the addition of Na considerably increases NAD levels, which do not show feedback inhibition effect on the enzyme (1). Here we describe the enzymatic characterization of human NaPRT. The protein was expressed in E. coli BL21(DE3) cells and purified to homogeneity by Ni-NTA resin affinity chromatography. The principal kinetic parameters of human NaPRT were determined. It was found that the enzyme followed a Michaelis-Menten kinetic with a random bireactant mechanism, and the Km for Na and PRPP were 44 μM and 22 μM, respectively. The Vmax was attested to be 0.013 U/mg, although in presence of Pi this value increased to 0.3 U/mg. Interestingly, ATP had a stimulation effect on enzymatic activity only at low substrates concentration, whereas at higher substrates concentration (1 mM), it showed an inhibitory effect. In addition to the kinetic characterization, the effect of several metabolites on NaPRT activity was investigated. As expected, NAD and NaAD, in addition to NMN and NaMN, had no effect on the enzyme activity, whereas piruvate and dihydroxyacetone phosphate showed a stimulation effect. On the contrary, several metabolites involved in the glucose or fatty acid metabolism significantly inhibited the enzyme, leading to hypothesize an in vivo NaPRT-depending regulation of NAD synthesis by these compounds. In addition, site-directed mutagenesis experiments were performed to elucidate the role of highly conserved residues. We determined that mutations, which involved residues localized in the / barrel domain of the protein, sensibly lowered the activity of the enzyme with respect of the wild-type protein. Human NaPRT was co-crystallized in presence of the potent enzyme inhibitor Acetyl-CoA using PEG 4000 as the major precipitant. The resulting crystals showed diffraction up to 3.0 Å resolution using synchrotron radiation at the ESRF, Grenoble France and are currently being optimised to undertake crystal structure determination. Finally, the 3D structure of the enzyme was predicted by homology modeling and used to carry out molecular docking simulations in order to identify the residues involved in the recognition and stabilization of ligands
Ruggeri, Barbara. "Espressione batterica e caratterizzazione dei determinanti strutturali dell'attività biologica della proteina PcF da Phytophthora cactorum." Doctoral thesis, Università Politecnica delle Marche, 2009. http://hdl.handle.net/11566/242365.
Повний текст джерелаMELONI, CLAUDIA. "Caratterizzazione strutturale e funzionale dei sistemi emoglobinici di due specie di pesci (Mugil cephalus e Ophisurus serpens) e di due varianti emoglobiniche umane (HbRoma e HbF-SS-Monserrato)." Doctoral thesis, Università degli Studi di Cagliari, 2012. http://hdl.handle.net/11584/266179.
Повний текст джерелаMojumdar, Aditya. "Structural and Biochemical study of human RECQ4." Doctoral thesis, Università degli studi di Trieste, 2015. http://hdl.handle.net/10077/11141.
Повний текст джерелаRecQ helicases belong to a ubiquitous family of DNA unwinding enzymes that are essential to maintain genome stability by acting at the interface between DNA replication, recombination and repair. Humans have five different paralogues of RecQ helicases namely RecQ1, BLM, WRN, RecQ4 and RecQ5. This work focuses on the structural and biochemical study of human RecQ4. Germ-line mutations in the RECQ4 gene give rise to three distinct human genetic disorders (Rothmund-Thomson, RAPADILINO and Baller-Gerold syndromes). Despite the important roles of RecQ4 in various cellular processes, RecQ4 have never been fully characterized. In addition to the helicase domain, RecQ4 has a unique N-terminal part that is essential for viability and is constituted by a region homologous to the yeast Sld2 replication initiation factor, followed by a cysteine-rich region, predicted to fold as a Zn knuckle. A part of this work focuses on the structural and biochemical analysis of both the human and Xenopus RecQ4 cysteine-rich regions, and shows by NMR spectroscopy that the Xenopus fragment does indeed assumes the canonical Zn knuckle fold, whereas the human sequence remains unstructured, consistent with the mutation of one of the Zn ligands. Both the human and Xenopus Zn knuckles bind to a variety of nucleic acid substrates, with a preference for RNA. We also investigated the effect of an additional Sld2 homologous region upstream the Zn knuckle. In both the human and Xenopus system, the presence of this region strongly enhances binding to nucleic acids. These results reveal novel possible roles of RecQ4 in DNA replication and genome stability. Recently the catalytic core of RecQ4 has been predicted to include RecQ-like-C-terminal (RQC) domain at the C-terminus of the helicase domain, similar to other RecQ helicases. This domain is composed of a Zn-binding region and a winged helix (WH) domain. Another part of this thesis centers on the structural and biochemical characterization of the catalytic core of RecQ4 including the helicase and RQC domain. The results provide an insight in the Zn binding ligands present in the RQC domain that plays a role in DNA binding and unwinding activity of the protein. Also the presence of the characteristic aromatic residue at the tip of the WH β hairpin and its role in DNA binding and unwinding has been established. Finally, it provides a low resolution SAXS model of the catalytic core of RecQ4.
Elicasi RecQ appartengono a una famiglia ubiquitaria di DNA svolgimento enzimi che sono essenziali per mantenere la stabilità del genoma agendo all'interfaccia tra replicazione del DNA, ricombinazione e riparazione. Gli esseri umani hanno cinque diversi paralogues di RecQ elicasi cioè RecQ1, BLM, WRN, RecQ4 e RecQ5. Questo lavoro si concentra sullo studio strutturale e biochimica di RecQ4 umana. Mutazioni germinali nel gene RECQ4 danno luogo a tre malattie genetiche umane distinte (Rothmund-Thomson, RAPADILINO e sindromi Baller-Gerold). Nonostante i ruoli importanti di RecQ4 in diversi processi cellulari, RecQ4 non sono mai stati pienamente caratterizzato. In aggiunta al dominio elicasi, RecQ4 ha una parte unica N-terminale che è essenziale per la vitalità ed è costituito da una regione omologa al lievito Sld2 fattore di iniziazione replica, seguita da una regione ricca di cisteina, previsto per piegare come stinco Zn . Una parte di questo lavoro si concentra sull'analisi strutturale e biochimica sia della regioni ricche di cisteina Xenopus RecQ4 umana e, e spettacoli di spettroscopia NMR che il frammento Xenopus effettivamente assume la canonica Zn nocca volte, mentre la sequenza di resti umani non strutturato, coerente con la mutazione di uno dei ligandi Zn. Sia il nocche Xenopus Zn umana e si legano ad una varietà di substrati di acido nucleico, con una preferenza per l'RNA. Abbiamo anche studiato l'effetto di un ulteriore regione omologa Sld2 monte la nocca Zn. Sia il sistema Xenopus umano e, la presenza di questa regione migliora fortemente legame ad acidi nucleici. Questi risultati rivelano possibili ruoli nuovi di RecQ4 nella replicazione del DNA e la stabilità del genoma. Recentemente il nucleo catalitico di RecQ4 stato previsto per includere RecQ-like-C-terminale (RQC) dominio al C-terminale del dominio elicasi, simile ad altri elicasi RecQ. Questo dominio è costituito da una regione-Zn vincolanti e un'elica alato (WH) dominio. Un'altra parte di questa tesi incentrata sulla caratterizzazione strutturale e biochimica del nucleo catalitico della RecQ4 compreso il elicasi e il dominio RQC. I risultati forniscono una descrizione nel Zn ligandi presenti nel dominio RQC che svolge un ruolo nel legame al DNA e l'attività svolgimento della proteina legante. Inoltre è stata stabilita la presenza della caratteristica residuo aromatico sulla punta della forcella WH β e il suo ruolo nel legame al DNA e di svolgimento. Infine, esso fornisce una bassa risoluzione SAXS modello del nucleo catalitico di RecQ4.
XXVII Ciclo
1985
Faccioli, Marco <1979>. "Organizzazione strutturale della catena respiratoria mitocondriale." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2010. http://amsdottorato.unibo.it/2886/1/Faccioli_Marco_Tesi_PDF.pdf.
Повний текст джерелаFaccioli, Marco <1979>. "Organizzazione strutturale della catena respiratoria mitocondriale." Doctoral thesis, Alma Mater Studiorum - Università di Bologna, 2010. http://amsdottorato.unibo.it/2886/.
Повний текст джерелаТези доповідей конференцій з теми "Caratterizzazione strutturale e biochimica"
Tognetti, R., S. Ravera, Bruno Lasserre, Ugo Chiavetta, M. Maesano, F. Lombardi, and Marco Marchetti. "Caratterizzazione strutturale e sink di carbonio in alcuni boschi vetusti e popolamenti persistenti d'Italia." In Terzo Congresso Nazionale di Selvicoltura. Accademia Italiana di Scienze Forestali, 2009. http://dx.doi.org/10.4129/cns2008.040.
Повний текст джерела