Interaction preferences in protein - DNA complexes Dávid Jakubec Abstract Interactions of proteins with DNA lie at the basis of many fundamental bio- logical processes. Despite ongoing efforts, the rules governing the recognition of specific nucleic acid sequences have still not been universally elucidated. In this work, I attempt to explore the recognition process by splitting the intricate network of contacts at the protein - DNA interface into contribu- tions of individual amino acid - nucleotide pairs. These pairs are extracted from existing high-resolution structures of protein - DNA complexes and in- vestigated by bioinformatics and computational-chemistry based methods. Criteria of specificity based on the coupling of observed geometrical prefer- ences and the respective interaction energies are introduced. The application of these criteria is used to expand the library of amino acid - nucleotide pairs potentially significant for direct sequence recognition. Electrostatic poten- tial maps are calculated for individual nucleotides as well as for selected complexes to investigate the physical basis of the observed specificity. 1

L'auto-organisation des organismes vivants est d'une complexité et d'une efficacité étonnantes. Plus précisément, les systèmes biologiques abritent un nombre gigantesque de réactions très spécifiques qui nécessitent que la bonne biomolécule se retrouve à la bonne place, dans le bon ordre et en un temps suffisamment court pour permettre le fonctionnement cellulaire, et au-delà la vie cellulaire. D'un point de vue dynamique, cela pose la question fondamentale de savoir comment les biomolécules trouvent efficacement leur(s) cible(s) spécifique(s), ou encore, quels types de forces rassemblent tous ces partenaires de réaction spécifiques dans un environnement aussi dense et ionisé que les micro-environnements cellulaires. Dans cette thèse, nous explorons la possibilité que des biomolécules puissent interagir à travers des interactions électromagnétiques de longue-portée telles que ces dernières sont prédites à partir des premiers principes de la physique; ''longue-portée'' signifiant que les interactionsen question sont actives sur des distances bien plus larges que les dimensions typiques des molécules mises en jeu (i.e., plus grandes qu'environ 50 angströms dans les systèmes biologiques). Après avoir posé les fondements théoriques concernant les interactionsde longue-portée potentiellement actives sur de longue distances dans un contexte biologique, nous étudions la posssibilité de détecter leur éventuelle contribution à partir de dispositifs expérimentaux qui sont accessibles de nos jours. Sur ce dernier point, des résultats préliminaires encourageants tant sur le plan théorique qu'expérimental sont présentés. / Self-organization of living organisms is of an astonishing complexity and efficiency. More specifically, biological systems are the site of a huge number of very specific reactions thatrequire the right biomolecule to be at the right place, in the right order and in a reasonably short time to sustain cellular function and ultimately cellular life. From the dynamic point of view, this raises the fundamental question of how biomolecules effectively find their target(s); in other words, what kinds of forces bring all these specific cognate partners together in an environment as dense and ionized as cellular micro-environments. In the present thesis, we explore the possibility that biomolecules interact through long-range electromagnetic interactions as they are predicted from the first principles of physics; "long-range" meaning that the mentioned interactions are effective over distances much larger than the typical dimensions of the molecules involved (i.e., larger than about 50 angströms in biological systems).After laying the theoretical foundations about interactions that are potentially active over long distances in a biological context, we investigate the possibility of detecting their contribution from experimental devices which are nowadays available. On the latter point, encouraging preliminary results both at the theoretical and experimental levels are exposed.

Dans les milieux naturels, la matière solide est essentiellement présente sous la forme de (bio)particules molles perméables aux ions et aux flux hydriques. Ces particules sont sans cesse soumises à des perturbations électriques/mécaniques, de telle sorte que les propriétés physico-chimiques des (bio)interphases qu'elles forment avec le milieu évoluent continûment dans le temps. Les (bio)interphases ne sont donc pas nécessairement à l'équilibre durant les processus interfaciaux (interactions électrostatiques, complexation de métaux). Dans ce contexte, nous avons évalué théoriquement l'énergie d'interaction électrostatique à l'équilibre entre (bio)particules molles multicouches de tailles et de densités de charge arbitraires. Puis nous avons déterminé l'impact de la dynamique hors-équilibre des propriétés électriques de (bio)films mous ligands sur leur capacité à former des complexes avec des métaux. Le dernier modèle théorique élaboré a pour objectif l'analyse de la dynamique de (bio)interphases multicouches hétérogènes en régimes d'équilibre et hors-équilibre. Enfin nous avons analysé à l'équilibre, en alliant l'AFM et l'électrophorèse, les propriétés mécaniques et électriques de bactéries E. coli exprimant spécifiquement (ou non) des structures de surface différentes. Toutes ces études ont montré la nécessité de prendre en compte pour l'analyse de la réactivité de (bio)particules dans leur milieu environnant (i) une représentation fidèle des (bio)particules (mollesse mécanique et hydrodynamique, hétérogénéité spatiale de la structure molle) et (ii) l'impact de la dynamique spatio-temporelle des (bio)interphases sur les processus gouvernant leur réactivité / In natural media, the solid matter is mainly present as soft (bio)particles (bacteria, viruses, humic acids) which are permeable toward ions and hydric fluxes. These (bio)particles are unceasingly exposed to electrical/mechanical perturbations, so that the physicochemical properties of (bio)interphases, developed by (bio)particles with the medium, evolve continuously. (Bio)interphases are thus not necessarily at equilibrium during interfacial processes e.g. electrostatic interactions, complexation with metallic contaminants. Under such a context, we evaluated theoretically at equilibrium the electrostatic interaction energy between soft multilayered (bio)particles with arbitrary sizes and charge densities. We then determined the impact of non- equilibrium electric properties of soft ligand polymeric (bio)films on their ability to form complexes with metals. The aim of the last theoretical model developed here is to analyze the dynamics of multilayered heterogeneous (bio)interphases in both equilibrium and non-equilibrium regimes. Finally we analyzed at equilibrium, by coupling AFM and microelectrophoresis measurements, mechanical and electrical properties of bacterial strains Escherichia coli that specifically express (or not) different surface structures (pili, fimbriae, adhesin Ag43). All these studies highlighted the necessity to integrate for the analysis of (bio)particles reactivity with their surrounding medium (i) a close representation of soft (bio)particles (mechanical and hydrodynamic softness, spatial heterogeneity of the soft material) and (ii) the impact of spatiotemporal dynamics of (bio)interphases on the processes governing their reactivity

Grâce à leurs propriétés physiques/chimiques uniques, les nanoparticules colloïdales sont au cœur de nombreuses applications innovantes. Afin de faciliter leur caractérisation ou de les intégrer dans des dispositifs fonctionnels, il est nécessaire de les assembler de manière dirigée sur des surfaces solides. Dans ce contexte, l’objectif de cette thèse est de mieux comprendre et d’optimiser la technique de nanoxérographie, méthode d’assemblage dirigé où des nanoparticules sont piégées sur des motifs de charges électrostatiques. Après un premier travail consistant à améliorer le procédé de nanoxérographie, trois problématiques spécifiques ont été adressées : (i) l’assemblage de particules micrométriques. Le couplage de simulations numériques et de manipulations expérimentales a permis d’identifier les paramètres clés de l’assemblage de telles particules colloïdales et d’élargir (facteur 100) la gamme de tailles de particules assemblables par nanoxérographie. (ii) l’analyse de l’assemblage multicouche. Par le biais de nanoparticules modèles luminescentes et par la mise en place d’un nouveau protocole d’assemblage, les critères clés génériques pour l’assemblage 3D de colloïdes par nanoxérographie ont été dégagés. (ii) l’assemblage dirigé de nanogels sensibles à un stimulus environnemental extérieur. L’utilisation d’un protocole d’assemblage optimisé a permis d’élaborer des assemblages de nanogels interactifs avec leur environnement et du faire du tri sélectif de ces nanoparticules sur une même surface. / Owing to their unique physico-chemical properties, colloidal nanoparticles are building blocks for the creation of plentiful innovative devices. In order to make easier their characterization and to incorporate them into functional nano-devices, it is necessary to perfectly control their directed assemblies onto solid surfaces. In this context, this thesis’ purpose is to simultaneously better understand and optimize the nanoxerography method, which allows electrostatic and selective directing assemblies of nanoparticles onto charged patterns. After an optimization of the nanoxerography process, three specific problematics have been addressed: (1) micron-sized particles assembly. The combined use of numerical simulations and experiments enabled to unveil the key parameters involved in micron-sized particles assembly and to expend the particle size range foreseeable for an assembly by nanoxerography (factor 100). (2) the 3D assembly analysis. The influence of diverse parameters on the 3D assembly of luminescent model nanoparticles was quantified by using a new assembly protocol. The results gave the generic key criterions for the 3D assembly of colloids by nanoxerography. (3) directed assembly of nanogels sensitive to an external environmental stimulus. The use of an optimized protocol allowed elaborating nanogels assemblies interactive with their environment and to sort these nanoparticles onto the same surface.

O consenso sobre os efeitos prejudiciais da material particulado atmosférico (MP) para a saúde humana é apoiado por um grande número de trabalhos epidemiológicos e experimentais mas a capacidade para definir qual o componente mais importante envolvido em sua toxicidade é dificultado pela sua grande complexidade de composição. O objetivo deste estudo é avaliar os efeitos de diferentes partículas de poluição atmosférica, quanto à origem e quanto a sua composição, sobre o sistema biológico através de 3 experimentos. O efeito do PM2,5 urbano de São Paulo, Brasil, coletado durante greve de ônibus foi avaliada através do TRAD-MN para a mutagenicidade e o teste de inibição de mitose em células meristemáticas da raiz de Allium cepa. para a toxicidade. Utilizamos o palato de rã para avaliar os parâmetros funcionais (transporte mucociliar -TMC, freqüência do batimento ciliar - FBC e diferença de potencial - DP) e morfométricos (espessura do epitélio, produção de muco) do aparelho mucociliar exposto ao ROFA, proveniente de usina siderúrgica e ao PM2,5 urbano. A análise comparativa da composição do PM2,5 através da ativação com nêutrons, fluorescência de raio-X e cromatografia gasosa espectrometria de massa mostrou redução na concentração de elementos traços de metais, redução de enxofre (44,17%) e de orgânicos (39,3%) na amostra de PM2,5 nos filtros coletados no dia de greve que associou-se com redução do dano no DNA. O ROFA induziu dano funcional do aparelho mucociliar (queda do TMC e aumento da FBC) que foi recuperado pela ação de antioxidantes vitamina E e n-Propil galato. O PM2,5 urbano provocou danos funcionais (diminuição do TMC e aumento da FBC) e morfológicos (diminuição da espessura e produção de muco) do epitélio respiratório. A análise comparativa da composição do ROFA e do PM2,5, demonstrou que o ROFA possui composição predominante de ferro (57,729%±0,004%) enquanto o PM2,5 urbano possui composição mais heterogênea, com predominância de enxofre (37,594%±17,930%), ferro (14,490%±6,489%) e silício (13,142%±8,516%) e pequenas quantidade de níquel (0,0336%±0,312%) e vanádio(0,260%±0,006%). Concluímos que substâncias oxidantes presentes no MP possuem papel relevante na injúria do aparelho mucociliar e que os efeitos lesivos do MP sobre os sistemas biológicos são fortemente dependentes de sua composição. / The association between atmospheric particulate matter (PM) and adverse human health effects have been consistently reported in epidemiological and experimental studies however, the ability to define the most important components involved in PM toxicity is hindered by its complex composition. In the present study we decided to further explore the effects of different atmospheric particles due to both source and composition, on the biologic system through 3 experiments. The mutagenicity and toxicity of urban PM2,5 of Sao Paulo, Brazil, from a bus strike day was evaluated by the TRAD-MN assay and inhibition of mitotic rate test of root tips Allium cepa. The frog palate was used to evaluate functional parameters (mucociliary transport ? MCT, ciliary beating frequency - CBF and potential difference ? PD) and morphometric data (epithelium tikness and mucus production) of the mucocliary apparatus exposed to ROFA from an a still plant, and to urban PM2,5. The comparative analysis of PM2,5 composition made by neutrons activation, fluorescence X-ray and gas cromatography/mass spectrometry showed decrease of trace elements, sulfur concentration (44,17%) and organics compounds (39,3%) in relation to the non strike day and was associated to coherent reduction in DNA damage. ROFA induced functional injury of the mucociliary apparatus (decrease of MCT and icrease of CBF) and the presence of antioxidants n-Propyl gallate and E vitamin protected against the mucociliary impairment induced by ROFA exposure. The comparative analysis between ROFA and PM2,5 composition showed that ROFA exhibited predominance of iron while PM2,5 showed heterogeneous composition presenting predominance of sulfur (37,594%±17,930%), iron (14,490%±6,489%) and silicon (13,142%±8,516%) and decrement of nickel (0,0336%±0,312%) and vanadium (0,260%±0,006%). We concluded that PM plays an important role in respiratory injury through oxidative stress and that adverse effects of PM on the biologic systems are strongly dependent on its composition.

Estudos teóricos dos mecanismos moleculares responsáveis pela formação e estabilidade de complexos moleculares vêm ganhando relevância pelas possibilidades práticas que oferecem, por exemplo, na compreensão de diversas doenças e no desenho racional de fármacos. Neste projeto, nossa ênfase está no estudo de complexos de proteínas, extraídos do banco de dados de proteínas (PDB), onde desenvolvemos ferramentas computacionais as quais permitem efetuar análises em duas direções: 1) efetuar previsões básicas, através do emprego de propriedades eletrostáticas de proteínas, em diferentes condições e níveis preditivos e 2) realização de um conjunto de análises estatísticas, como freqüência de contato, em busca de preditores de complexos de proteínas e identificar padrões de interação entre seus aminoácidos em função da distância de separação. Com base nos resultados obtidos por ambos os estudos, objetivamos quantificar as forças físicas envolvidas na formação dos complexos protéicos. O foco do projeto, a longo prazo, é prever o fenômeno da complexação através da fusão dessas duas linhas de estudos: preditor básico de complexos protéicos e análise do potencial estatístico entre os aminoácidos que formam o complexo. O presente projeto é concluído com a construção de portais web que disponibilizarão os resultados obtidos por nossos trabalhos bem como a possibilidade de qualquer usuário, efetuar consultas por propriedades de proteínas e/ou grupo de proteínas. / Theoretical studies of the molecular mechanisms responsible for the formation and stability of molecular complexes are gaining relevance for the practical possibilities that they offer, for example, in the understanding of diverse diseases and rational drug design. In this project, our emphasis is on the study of protein complexes, extracted from protein data bank (PDB). We have developed computational tools which allow to perform analyses in two directions: 1) to make basic complexation forecasts, through the use of electrostatics properties of proteins, in different conditions and predictive levels, and 2) to carry out a set of statistical analyses, as contacts frequency, in order to build up predictor of protein complexes and to identify patters of interactions between the amino acids as a function of their separation distance. Based on the results obtained on both studies, we aim quantify the physical forces involved in the formation of protein complexes. The focus of the project, in the long run, is to foresee the phenomenon of the protein complexes through the fusing of these two study lines: a coarse-grained predictor of protein complexes and analysis of the statistical potentials between the amino acids that form the complex. The present project is concluded with the construction of web services where we make available the results obtained on our works. This server also has the possibility to be used by any computer user, that wishes to perform search on protein and/or protein group properties

Macromolecular biomaterials often require covalent crosslinking to achieve adequate stability and mechanical strength for their given application. However, the use of auxiliary chemicals may be associated with long-term toxicity in the body. Oppositely-charged polyelectrolytes (PEs) have the advantage that they can self-crosslink electrostatically and those derived from marine organisms are an inexpensive alternative to glycosaminoglycans present in the extracellular matrix of human tissues. A range of different combinations of PEs and preparation conditions have been reported in the literature. However, although there has been some work on complex formation between chitosan (CS) and carrageenan (CRG), much of the work undertaken has ignored the effect of pH on the consequent physicochemical properties of self-crosslinked polyelectrolyte complex (PEC) gels, films and scaffolds. Chitosan is a positively-charged polysaccharide with NH3+ side groups derived from shrimp shells and, carrageenan is a negatively-charged polysaccharide with OSO3- side groups derived from red seaweed. These abundant polysaccharides possess advantageous properties such as biodegradability and low toxicity. However, at present, there is no clear consensus on the cell binding properties of CS and CRG or CS-CRG PEC materials. The aim of this study was to explore the properties of crosslinker-free PEC gels, solvent-cast PEC films and freeze-dried PEC scaffolds based on CS and CRG precursors for medical applications. The objective was to characterise the effect of pH of the production conditions on the physicochemical and biological properties of CS-CRG PECs. Experimental work focused on the interaction between PEs, the composition of PECs, the rheological properties of PEC gels and the mechanical properties of PEC films and scaffolds. In addition, cell and protein attachment to the PEC films was assessed to determine their interactions in a biological environment. For biomedical applications, these materials should ideally be stable when produced such that they can be processed to form either a film or a scaffold and have mechanical properties comparable to those of collagenous soft tissues. FTIR was used to confirm PEC formation. Zeta potential measurements indicated that the PECs produced at pH 2-6 had a high strength of electrostatic interaction with the highest occurring at pH 4-5. This resulted in stronger intra-crosslinking in the PEC gels which led to the formation of higher yield, solid content, viscosity and fibre content in PEC gels. The weaker interaction at pH 7-12 resulted in higher levels of CS incorporated into the complex and the formation of inter-crosslinking through entanglements between PEC units. This resulted in the production of strong and stiff PEC films and scaffolds appropriate for soft tissue implants. The PECs prepared at pH 7.4 and 9 also exhibited low swelling and mass loss, which was thought to be due to the high CS content and entanglements. From the range of samples tested, the PECs produced at pH 7.4 appeared to show the optimum combination of yield, stability and homogeneity for soft tissue implants. Biological studies were performed on CS, CRG and PECs prepared at pH 3, 5, 7.4 and 9. All of the PE and PEC films were found to be non-cytotoxic. When the response of three different cell types and a high binding affinity protein (tropoelastin) was evaluated; it was found that the CS-CRG PEC films displayed anti-adhesive properties. Based on these experimental observations and previous studies, a mechanistic model of the anti-adhesive behaviour of PEC surfaces was proposed. It was therefore concluded that the CS-CRG PECs produced might be suitable for non-biofouling applications.

La démarche expérimentale menée dans cette thèse CIFRE a eu comme objectif l’étude de la faisabilité de la séparation électrostatique d’une large gamme de mélanges de matériaux granulaires issus de déchets d’équipements électriques et électroniques (conducteur – isolant, conducteur – conducteur et isolant – isolant). Trois mécanismes physiques de charge électriques ont été mis en oeuvre : l’induction électrostatique, la décharge couronne et l’effet triboélectrique. Un dispositif de charge de laboratoire et deux séparateurs électrostatiques industriels ont été conçus et réalisés. Le dispositif de charge à cylindre(s) tournant(s) permet la charge triboélectrique de matériaux plastiques. Le prototype de séparateur industriel a d’abord été réalisé pour le traitement de mélanges granulaires conducteur – isolant puis a été adapté à la séparation isolant – isolant. Les trois mécanismes de charge peuvent y être utilisés. Ce prototype a permis la fabrication du premier séparateur électrostatique de série de la société CITF dédié à la séparation de déchets de câbles électriques. Des mesures de charge et de masse ainsi que des simulations et visualisations des trajectoires des particules par caméra rapide ont facilité l’analyse des phénomènes. La méthode des plans d’expérience a permis d’identifier les facteurs influents sur les processus de séparation et de définir leur point de fonctionnement optimal. L’influence d’une décharge à barrière diélectrique sur la charge triboélectrique et la séparation électrostatique de mélanges plastiques a été étudiée. Les travaux réalisés dans cette thèse ont confirmé la séparabilité électrostatique de mélanges de matériaux granulaires. / The experimental approach used in this thesis was aimed at the study of the feasibility of the electrostatic separation of a wide range of granular mixtures from waste electric and electronic equipment (conductive – insulator, conductive – conductive, insulator – insulator). Three physical mechanisms of electrical charging have been employed: electrostatic induction, corona discharge and triboelectric effect. A tribocharging device and two industrial electrostatic separators have been designed and engineered. The rotating cylinder tribocharging device allows the triboelectric charging of plastics. The industrial separator prototype was firstly built for processing conducting – insulator granular mixtures and then adapted for insulator – insulator separation. This prototype, which enables the use of all three electrical charging mechanisms, served as model for the CITF company first industrial electrostatic separator for the recycling of metals and plastics from electric cable wastes. Charge and mass measurement, as well as modelling and visualization of particle trajectories by a high-speed camera facilitated the investigations. Design of experiments method was used to define the influential factors and find the optimum operating conditions of the separation processes. Dielectric barrier discharge exposure of granular plastics may enhance the efficiency of the triboelectric charging of these materials and hence facilitate their electrostatic separation. These researches have confirmed the feasibility of the electrostatic separation of a wide variety of granular mixtures.

No presente trabalho é apresentado um conjunto de medidas de seção de choque do processo de fusão nuclear nos sistemas ANTPOT. 12 C + ANTPOT. 63, 65 CU, em um intervalo de energia correspondente à 0.91.8 vezes a barreira coulombiana. O método experimental utilizado foi a detecção dos resíduos de evaporação através da técnica de tempo de vôo, acoplado à um defletor eletrostático para separar as partículas provenientes do espalhamento elástico dos produtos de fusão. As vantagens e limitações do método são discutidas em detalhes. As funções de excitação de fusão dos sistemas citados foram analisadas via modelo de penetração de barreira unidimensional , utilizando vários potenciais nucleares. Dessa análise comprovou-se que a secção de choque de fusão é subestimada em energias em torno e abaixo da barreira coulombiana. Nesse sentido foi feito um cálculo, utilizando o código computacional CCFUS, que simula o acoplamento de canais, com a finalidade de descobrir quais canais contribuem para o processo de fusão nuclear em baixas energias. As seções de choque de fusão obtidas foram comparadas com outros sistemas - ANTPOT. 12 C + ANTPOT. 63, 65 CU.- que utilizam os mesmos núcleos-alvo. A comparação revelou que os sistemas ANTPOT. 12 C + ANTPOT. 63, 65 CU possuem o maior aumento de seção de choque de fusão subcoulombiana e esse fato pode estar relacionado à deformação estática do projétil. A comparação das distribuições da velocidade dos resíduos de evaporação para os dois de sistemas revelou um canal intenso para o sistema ANTPOT. 12 C + ANTPOT. 63 CU que não está presente para o outro sistema. / In this work we present cross-section measurements for nuclear fusion in the 12C+ 63,65 Cu system, at 12C energy range from 0.9 to 1.8 times the Coulomb barrier. In order to detect and to obtain the mass identification of the evaporation residues following the fusion process, the time of flight method was adopted in conjunction with an eletrostatic deflector capable of separating the evaporation residues from the beam particles. The limitation and advantadges of this method of measurement are discussed. The excitation functions for the above systems were analysed using the unidimensional barrier penetration model with different nuclear potentials. Theoretical fusion cross-section values obtained from this analysis were systematically smaller than our measured values, in the energy region below the Coulomb barrier. In order to discover which channel enhances the fusion cross-section in this region, a coupled channel calculation was performed, with the aid of CCFUS code. The experimental data for the above reactions were compared with the systems 12C+ 63,65 Cu, measured by our group. In this comparison, It was noted that the systems 12C+ 63,65 Cu have greater fusion cross section below the Coulomb barrier. This fact can be related to the static deformation of 12C. The comparison of velocity spectra of the evaporated residues for the two systems shows that 12C+63Cu has a strong reaction channel that was not present in the 12C+ 65Cu system.

No presente trabalho é apresentado um conjunto de medidas de seção de choque do processo de fusão nuclear nos sistemas ANTPOT. 12 C + ANTPOT. 63, 65 CU, em um intervalo de energia correspondente à 0.91.8 vezes a barreira coulombiana. O método experimental utilizado foi a detecção dos resíduos de evaporação através da técnica de tempo de vôo, acoplado à um defletor eletrostático para separar as partículas provenientes do espalhamento elástico dos produtos de fusão. As vantagens e limitações do método são discutidas em detalhes. As funções de excitação de fusão dos sistemas citados foram analisadas via modelo de penetração de barreira unidimensional , utilizando vários potenciais nucleares. Dessa análise comprovou-se que a secção de choque de fusão é subestimada em energias em torno e abaixo da barreira coulombiana. Nesse sentido foi feito um cálculo, utilizando o código computacional CCFUS, que simula o acoplamento de canais, com a finalidade de descobrir quais canais contribuem para o processo de fusão nuclear em baixas energias. As seções de choque de fusão obtidas foram comparadas com outros sistemas - ANTPOT. 12 C + ANTPOT. 63, 65 CU.- que utilizam os mesmos núcleos-alvo. A comparação revelou que os sistemas ANTPOT. 12 C + ANTPOT. 63, 65 CU possuem o maior aumento de seção de choque de fusão subcoulombiana e esse fato pode estar relacionado à deformação estática do projétil. A comparação das distribuições da velocidade dos resíduos de evaporação para os dois de sistemas revelou um canal intenso para o sistema ANTPOT. 12 C + ANTPOT. 63 CU que não está presente para o outro sistema. / In this work we present cross-section measurements for nuclear fusion in the 12C+ 63,65 Cu system, at 12C energy range from 0.9 to 1.8 times the Coulomb barrier. In order to detect and to obtain the mass identification of the evaporation residues following the fusion process, the time of flight method was adopted in conjunction with an eletrostatic deflector capable of separating the evaporation residues from the beam particles. The limitation and advantadges of this method of measurement are discussed. The excitation functions for the above systems were analysed using the unidimensional barrier penetration model with different nuclear potentials. Theoretical fusion cross-section values obtained from this analysis were systematically smaller than our measured values, in the energy region below the Coulomb barrier. In order to discover which channel enhances the fusion cross-section in this region, a coupled channel calculation was performed, with the aid of CCFUS code. The experimental data for the above reactions were compared with the systems 12C+ 63,65 Cu, measured by our group. In this comparison, It was noted that the systems 12C+ 63,65 Cu have greater fusion cross section below the Coulomb barrier. This fact can be related to the static deformation of 12C. The comparison of velocity spectra of the evaporated residues for the two systems shows that 12C+63Cu has a strong reaction channel that was not present in the 12C+ 65Cu system.