Symmetric cell division in gram-negative bacteria is essential for generating two equal-sized daughter cells, each containing cellular material crucial for growth and future replication. The Min system, comprised of proteins MinC, MinD and MinE, is particularly important for this process since its deletion leads to minicells incapable of further replication. This thesis focuses on the interactions involving MinE that are important for allowing cell division at the mid-cell and for directing the dynamic localization of MinD that is observed in vivo. Previous experiments have shown that the MinE protein contains an N-terminal region that is required to stimulate MinD-catalyzed ATP hydrolysis in the Min protein interaction cycle. However, MinD-binding residues in MinE identified by in vitro MinD ATPase assays were subsequently found to be buried in the hydrophobic dimeric interface in the MinE structure, raising the possibility that these residues are not directly involved in the interaction. To address this issue, the ability of N-terminal MinE peptides to stimulate MinD activity was studied to determine the role of these residues in MinD activation. Our results implied that MinE likely undergoes a change in conformation or oligomerization state before binding MinD. In addition we performed circular dichroism spectroscopy of MinE. The data suggest that direct interactions between MinE and the lipid membrane can lead to conformational changes in MinE. Using NMR spectroscopy in an attempt to observe this structure change, different membrane-mimetic environments were tested. However the results strongly suggest that structural studies on the membrane-bound state of MinE will pose significant challenges. Taken together, the results in this thesis open the door for further exploration of the interactions involving MinE in order to gain a better understanding of the dynamic localization patterns formed by these proteins in vivo.

De nombreux travaux ont été dédiés ces dernières années aux matériaux à transition de spin qui présentent des potentialités pour le stockage de l’information en raison de leurs propriétés physiques commutables : métastabilité thermique et optique. Ce travail est relatif au mécanisme photo-induit dans les systèmes à transition de spin et, plus précisément, à l’étude des effets de dilution sur la température LIESST. L’objectif principal est de mieux cerner les paramètres qui gouvernent T(LIESST). Diverses études expérimentales ont été réalisées sur des composés mononucléaires 0D, polymériques 1D et polynucléaires. L’analyse de l’ensemble des propriétés photomagnétiques a permis de montrer que T(LIESST) est indépendant de la dilution et suggère que cette grandeur est gérée par la molécule et dépend très peu des interactions intermoléculaires. / In the last few years, many studies have been dedicated to spin-crossover compounds because of their potential application in data-storage systems. In such systems, optical switching is particularly promising. This work deals with the photo-induced phenomenon encountered in spin crossover systems, more specifically, the effects of metal dilution on the LIESST temperature. The main objective is to better define the parameters that govern T (LIESST). Various experimental studies were performed on mononuclear 0D compounds, polymeric 1D systems and polynuclear systems. The analysis of all photomagnetic properties has shown that T(LIESST) is not affected by the dilution and suggests that this parameter is govern at the molecular scale and depends very few of intermolecular interactions.

O estudo das reações de agregação em sistemas automontados é de grande interesse científico pela grande variedade de aplicações que tais colunas podem desempenhar. O presente trabalho teve como objetivo investigar as propriedades gerais de colunas supramoleculares formadas por complexos de coordenação que contêm platina, paládio e zinco. Esta investigação foi feita através do método semiempírico PM7. Os agregados supramoleculares investigados tiveram suas geometrias otimizadas e foram submetidos a cálculos termoquímicos. Pôde-se investigar as influências do metal e dos ligantes nos mecanismos gerais e propriedades das colunas, uma vez que utilizaram-se dois ligantes distintos. Estudou-se compostos de coordenação cujos ligantes foram o 2,6-bis(1H-1,2,4-triazol-5-il)piridina e oligofenilenoetinilenos (OPE) piridínicos. Realizou-se cálculos utilizando o modelo de solvatação contínua COSMO com água e metilciclohexano (MCH), o que permitiu avaliar os efeitos dos solventes nas energias de dimerização dos sistemas estudados. Os resultados obtidos para os derivados de OPE indicaram que as colunas supramoleculares desta classe que contêm Zn(II) apresentam anticooperatividade, exibindo energias de Gibbs de formação para o vácuo a 298 K positivas do dímero ao decâmero. Os resultados de solvatação contínua indicaram que a formação de um dímero deste composto é termodinamicamente favorável de 278 K para água e 267 K para o MCH, enquanto que para o vácuo a temperatura na qual a energia de Gibbs de dimerização é negativa ocorre em 246 K. Tal resultado aponta que o principal efeito de formação de colunas baseadas neste composto em altas temperaturas é primariamente um efeito solvofóbico que depende da polaridade do solvente. Observou-se a cooperatividade em vácuo e em ambos os solventes das colunas de derivados de OPE que contêm Pd(II) e Pt(II). Além destes fatos, as distâncias entre os centros metálicos diminuíram conforme monômeros eram adicionados à coluna, o que indica a existência de interações metalofílicas no sistema. Este resultado é corroborado experimentalmente por um artigo de Albuquerque el al. no caso do sistema de derivado de OPE que contém Pd(II). Para os sistemas cujos ligantes eram a 2,6-bis(1H-1,2,4-triazol-5-il)piridina, houve cooperatividade para ambas as colunas baseadas em Pd(II) e Pt(II), e houve para o sistema cujos centros metálicos são o Pt(II) uma transição não observada para todos os outros sistemas estudados; houve comportamento distinto das mudanças de distâncias entre monômeros no sistema triazol pridínico baseado em Pd(II) em relação àquele baseado em Pd(II) derivado de OPE. Este resultado indica a soma de dois efeitos no sistema OPE baseado em Pd(II): o de interações metalofílicas e de empilhamentos π-π. Como um todo, os resultados indicam que as principais interações envolvidas nas formações das colunas supramoleculares são interações de empilhamento π-π e metalofílicas para quatros compostos em graus diversos, exceto para o derivado de OPE baseado em Zn(II), que tem como principal força-motriz de agregação interações solvofóbicas. Perspectivas para este projeto: continuação da modelagem dos sistemas estudados por dinâmica molecular, com a simulação das interações de várias colunas formadas pelos monômeros estudados através do CP2K e publicação de um artigo com os resultados obtidos. / The study of aggregation reactions in self-assembled systems is of great scientific interest due to the wide range of applications which such resulting columns can play. Herein, we had the objective of investigating the general properties of supramolecular columns formed coordination complexes which contain platinum, palladium and zinc. This investigation was carried through the semiempirical method PM7. The investigated supramolecular aggregates were optimized, and their optimized geometries were subjected to thermochemical calculations. In this sense we could probe the influences of metals and ligands onto the general mechanisms and columnar properties, once were herein used two different ligands. The two ligands were 2,6-bis(1H-1,2,4-triazol-5-il)pyridine and oligophenyleneethynediyl (OPE) pyridine derivatives. COSMO solvation model -based calculations in water and methylcyclohexane (MCH) were performed in order to evaluate solvents effects on dimerization Gibbs energies of the studied systems. The results obtained for the OPE derivative based on Zn(II) pointed an anticooperative process in vacuum at 298K, exhibiting positive Gibbs energies of formation from the dimer to the decamer. The results from continuum solvation calculations indicate the formation of a dimer based on this Zn(II) molecule to be thermodynamically favorable at 278 K for water and at 267 K for MCH, while in vaccum this point lies at 246 K. This results indicates the formation of supramolecular columns based on this compound are formed in higher temperatures primarily due to solvophobic effects, and the intensity of this effect depends on the solvent polarity. The columns of OPE derviatives which contained Pd(II) and Pt(II) both showed cooperativity in the aggregation process. Besides these thermodynamic results, both of those columns showed monotonic distance shortening due to columns increasing process. This is an indicative of metallophilic interactions taking place in these aggregates. This result for Pd(II) is experimentally supported by a paper od Albuquerque et al. for the columns containing 2,6-bis(1H-1,2,4-triazol-5-il)pyridine cooperativity was observed for both the columns--those which contain Pd(II) and Pt(II)--, and for the Pt(II)-based columns of this class it was observed a transition not of most eneregetically favoured mechanisms not observed for all the other systems studied. A difference on the behavior of average monomer distances took place in the triazol pyridine Pd(II) compound when compared to its metal-related OPE derivative. This result indicates the summation of effects in the OPE-based Pd(II) system: the one of metallophilic interactions and π-π stacking interactions. As a whole, the results indicate π-π stacking and metallophilic interactions to be responsible for aggregation processes in diverse intensities for four of the compounds, except for that in the Zn(II)-based OPE derivative, which has the main aggregation effect as solvophobic interactions. The main perspectives for this project are the continuation of the modelling of the herein studied systems by molecular dynamics, simulating the interactions of many columns by the CP2K program, and the publication of a paper which contains the results obtained and presented in this dissertation.

O gênero Piper pertencente à família Piperaceae, encontra-se distribuído nas regiões tropicais e subtropicais do globo. Estudos químicos têm demonstrado diversidade de metabólitos secundários com atividade biológica. Os alcalóides são metabólitos característicos. A piplartina, (E)-1-(3-(3,4,5-trimetoxifenil)acriloil)-5,6- diidropiridin-2(1H)-ona, é um alcalóide encontrado em muitas espécies. Tem atividade citotóxica contra células de linhagem tumoral, ansiolítica, antidepressiva, antifúngica e antiagregação plaquetária, sendo dessa forma, uma molécula candidata a um novo fármaco. O conhecimento do metabolismo de um candidato a fármaco é um fator importante na avaliação da sua segurança e eficácia. Ensaios in vitro estão crescentemente sendo utilizados como screening e os microssomas hepáticos representam o sistema in vitro mais utilizado. Dessa forma, o presente trabalho tem como objetivo determinar os parâmetros cinéticos enzimáticos in vitro da piplartina utilizando microssomas de fígado de ratos, bem como a determinação dos possíveis metabólitos formados. Para tanto, foi desenvolvido um método de quantificação da piplartina utilizando cromatografia líquida de alta eficiência. Como condição de análise, empregou-se uma coluna C18, fase móvel acetonitrila:água (40:60, v/v) e vazão de 1 mL min-1. Para extração da piplartina dos microssomas hepático de ratos foi empregado a extração líquido-líquido utilizando 4,0 mL de hexano como solvente extrator. Após otimização da extração, o método foi validado, mostrando-se linear na faixa de 2,4-157,7 ?M, obtendo-se uma equação da reta y= 0,0934x + 0,0027, (r= 0,99) e limite de quantificação de 2,4 ?M. A recuperação média foi de 85%. A precisão e exatidão apresentaram resultados dentro do recomendável pela ANVISA. A piplartina manteve-se estável até 50 minutos em condições de incubação, e até 6h sob a bancada. Após validação da metodologia, estabeleceram-se as condições lineares para a quantidade de proteínas microssomais: 0,28 mg mL-1 e para o tempo de incubação: 16 minutos no consumo da piplartina no meio microssomal, e então efetuou-se a determinação dos parâmetros cinéticos enzimáticos da piplartina empregando as condições de V0. Nesse estudo foi observado um Vmax= 4,74 ± 0,26 ?M/?g mL-1/min, h= 2,53 ± 0,37, S50= 44,69 ± 0,32 ?M e CLmax= 0,054 ?L/min/mg proteina, um perfil cinético indicativo de cooperatividade. Um estudo qualitativo para determinação dos possíveis metabólitos foi feito utilizando-se a espectrometria de massas, por meio da qual foi possível identificar a formação de dois produtos hidroxilados. Deste modo, os microssomas mostraram-se uma ferramenta útil, rápida e simples para determinação da cinética enzimática, e na condução dos estudos preliminares de metabolismo in vitro. / The genus Piper belongs to the Piperaceae family and includes species that are widely distributed throughout the tropical and subtropical regions of the world. Chemical studies have shown diversity of secondary metabolites with biological activity. The alkaloids are characteristic metabolites. The piplartine, (E)-1-(3-(3,4,5- trimethoxyphenyl)acryloyl)-5,6-diidropiridin-2(1H)-one is an alkaloid found in many species. It shows cytotoxic activity against tumor cell lines, anxiolytic, antidepressant, antifungal, and antiplatelet therapy, thus being a drug candidate. The knowledge regarding the oxidative metabolism is an important tool in assessing the safety and efficacy of a drug candidate. In vitro assays are increasingly being used as a screening tool and liver microsomes represent the most widely in vitro system used for that. This study aims to determine the in vitro enzymatic kinetic parameters for piplartine by cytochrome P450 enzymes (CYP) present in the rat liver microsomes, and the determination of possible metabolites. To accomplish, it was developed a method to quantify the piplartine using high performance liquid chromatography. The analysis was carried out employing a C18 column, mobile phase: acetonitrile: water (40:60, v/v) at a flow rate of 1 ml min-1. To extract piplartine from rat liver microsomes it was employed the liquid-liquid extraction (4.0 mL of hexane). The method was validated and proved to be linear in the range of 2.4 to 157.7 ?M, the equation for calibration curve was: y= 0.0934x + 0.0027 (r = 0.99), and a limit of quantification of 2.4 ?M. The mean recovery was 85%. The precision and accuracy were in agreement with ANVISA guidelines. The piplartine remained stable until 50 minutes of incubation conditions, and until 6 hours under the bench. Once validated, it was set the conditions for the linear amount of microsomal protein: 0.28 mg mL-1 and to the incubation time: 16 minutes, then it was performed the determination of enzymatic kinetic parameters, that revealed a sigmoidal profile with Vmax = 4.74 ± 0.26 ?M/mg mL-1/min, h = 2.53 ± 0.37, S50 = 44.69 ± 0.32 ?M, and CLmax = 0.054 ?L/min/mg protein, indicating a cooperativity behavior. A qualitative study to determine possible metabolites carried out using mass spectrometry, through which it was possible to identify the formation of two hydroxylated products. To conclude, the microsomes showed to be a useful, fast and simple tool to determination of enzymatic kinetics and in vitro metabolism studies.

Cette thèse traite des problèmes d’estimation et de contrôle de cohérence par l’utilisation des techniques ensemblistes. L’objectif est la mise en place d’une démarche méthodologique pour la surveillance et la détection d’anomalies au sein des systèmes où le déterminisme des indicateurs relatifs à l’état de santé du système est une condition sine qua non. Une fois placé dans un contexte ensembliste, l’évolution de chaque variable du système est représentée par une enveloppe traduisant les incertitudes internes et externes ; cette enveloppe représente le seuil au delà duquel le comportement observé représente un écart anormal par rapport à son comportement nominal, et pouvant conduire à une incapacité pour accomplir les objectifs de sa mission. Les techniques développées sont appliquées à un procédé hydraulique de laboratoire . / This work deals with the development of set-membership methods for set esti- mation and consistency checks for nonlinear continuous-time systems. The main objective is to setup a methodology for fault detection and isolation for the systems where the deter- minism of faults indicators on the health system is a necessary condition. Once placed in a set-membership framework, the evolution of each variable is represented by an envelope re?ecting the internal and external uncertainties. This envelope corresponds to the threshold beyond which the observed behavior is an abnormal discrepancy over its nominal behav- ior, thus preventing the accomplishment the mission objectives. The proposed methods are applied on a hydraulic laboratory process.

Ce travail de thèse porte sur l'étude de la commutation ultrarapide de matériaux moléculaire photomagnétiques présentant des transitions entre états de spin. Ces cristaux moléculaires sont des prototypes de bi-stabilité moléculaire possédant deux états électroniques distincts, Haut Spin (HS) et Bas Spin (LS), entre lesquels les molécules peuvent commuter par la lumière. L'émergence des techniques ultrarapides nous permet d'étudier en temps réel ces processus de photo-commutation ainsi que les dynamiques hors équilibres associées jusqu'à l'échelle femtoseconde (10-15s). Nous avons combiné ici l'utilisation de sondes sensibles aux changements d'états électroniques et aux changements structuraux pour étudier ces processus de photo-commutation. Des mesures d'absorption optiques femtosecondes ont été effectuées sur notre plate forme laser et elles ont complétées par des mesures de diffraction et d'absorption des rayons X résolues en temps. La première partie de cette thèse se focalise sur la dynamique de commutation induite par la lumière au niveau moléculaire. Elle révélé l'intrication compliquée de degrés de liberté électroniques et structuraux. La génération et l'amortissement rapide de phonons optiques est identifié comme étant le processus clé dans le piégeage des molécules dans l'état Haut Spin. Les mesures réalisées sur différents types de composés ont prouvé le caractère local et linéaire de ce processus. La seconde partie de cette thèse présente les études de dynamique hors équilibre et des effets en cascade d'origine élastique et thermique résultant de ces perturbations initiales. Des effets coopératifs induits par la lumière sont ainsi mis en évidence. Cette dynamique hors équilibre pilotée par des phénomènes propagatifs et diffusifs est sensible aux effets de taille. L'étude de nano-cristaux démontre une grande efficacité et une réponse aux effets élastiques plus rapide que dans le cas des macrocristaux. Ces études apportent une compréhension nouvelle des phénomènes hors équilibre liés aux transitions de phase photo-induites sur des échelles de temps et d'espace allant de la molécule au matériau. / The main topic of this thesis is the study of the ultrafast photo-switching of photo-magnetic molecular materials showing transition between spin states. These molecular crystals are prototypes of molecular bistability between two distinct electronic states, HS and LS. The molecules can be switched between these two states by a light pulse. The emergence of ultrafast techniques allows us to study in real time these photo-switching processes and also the associated out-of-equilibrium dynamics down to the femtosecond scale (10-15s). We have combined probes sensitive to the change of electronic state, on the hand, and to structural rearrangements, on the other hand, in order to observe these photo-switching processes. The measurements of ultrafast transient absorption spectroscopy have been made using the laser plateform at the IPR. Complementary time resolved X-ray diffraction and absorption experiments have been performed on large facilities. The first part of this manuscript is focused on the photo-switching dynamics at molecular scale. It reveals a complicated interaction between electronic and structural degrees of freedom. The generation and damping of coherent optical phonons is identified as a key parameter in the trapping in HS potential. Several experiments on different compounds show the linear and local character of such ultrafast photo-switching. The second part of this thesis presents studies on the complete out of equilibrium dynamics. It reveals a cascading process with activation of elastic and thermal effects at different time scales. Cooperative processes following a light excitation are observed. These complexes dynamics are driven by propagating and diffusive process sensitive to the size of the sample. The study of nanocrystals yields high conversion and faster response to elastics effect than single-crystals. These studies further elucidate the out of equilibrium processes underlying the photo-induced phase transitions on time and length scales, from the molecule to material scale.

Ce travail de thèse porte sur la chimie organométallique des complexes pince indényle/indénediide de palladium et de platine et leurs applications en catalyse coopérative métal-ligand. Le premier chapitre de ce manuscrit fait un point bibliographique non-exhaustif du domaine de la coopérativité métal-ligand, des travaux de Noyori sur l'hydrogénation asymétrique, jusqu'aux récents travaux de Milstein avec les ligands désaromatisés. Les ligands pince sont également présentés. Du premier exemple PCP décrit par Shaw jusqu'au CNC développé par Bezuidenhout. La versatilité de ces ligands est illustrée à travers quelques modifications permettant des réactivités originales ou l'isolation d'espèces hautement instables. Le second chapitre présente une nouvelle réaction de formation de cycle avec le complexe pince indénediide de palladium. Cette réaction implique pour la première fois deux molécules de substrat, le CO2 comme source C1 et les propargylamines/homopropargylamines. L'étude mécanistique du système a permis d'établir l'implication de la coopérativité métal-ligand. Ensuite, le troisième chapitre est un chapitre de chimie exploratoire. De nouvelles réactivités ont été étudiées avec les complexes pince de platine. L'activation de liaisons peu polaires telles que H-H et H-Si ont permis la réduction de liaisons insaturées. Les propositions mécanistiques, qu'il reste à confirmer, semblent indiquer que les métathèses ?, ainsi que les insertions migratoires, sont possibles avec ces complexes. Finalement, le dernier chapitre est consacré au développement d'un nouveau ligand pince ayant la particularité d'être hémilabile et ouvrant ainsi la voie à de nouvelles réactivités. Sa coordination au palladium, ainsi que la déprotonation de ce dernier, a permis le développement d'un complexe coopératif qui a été testé en cycloisomérisation. L'ensemble de ces travaux reflètent l'importance des ligands pince indényle/indénediide et de la coopérativité métal-ligand en catalyse. / This Ph.D. work deals with organometallic chemistry of indenyl/indenediide palladium and platinum pincer complexes and their applications in metal-ligand cooperative catalysis. The first chapter of this manuscript compiled a non-exhaustive bibliographic survey of the field of metal-ligand cooperation, from Noyori's system applied to the asymmetric hydrogenation, to the recent examples described by Milstein involving non-aromatic pincer ligands. Pincer ligands are also presented. Starting from the first example, in which Shaw shed light a PCP pincer, to the contemporary CNC pincer reported by Bezuidenhout. The versatility of those ligands is illustrated through few modulations allowing originals reactivities or stabilisation of highly unstable species. The second chapter is focused on the development of a new catalytic reaction with the indenediide palladium pincer complex. This reaction entail, for the first time, two substrates, the CO2 as a C1 source and propargylamines/homopropargylamines. The mechanistic studies turn out the importance of the metal-ligand cooperativity. Then, the third chapter concerns exploratory chemistry. New reactivities have been studied with the platinum complexes. The activation of low polar bond such as H-H and H-Si allowed the reduction of unsatured C-C bond. The mechanistic propositions, that remain to be confirmed, seem to indicate the feasibility of ? bond metathesis and migratory insertions. Finally, the last chapter is dedicated to the development of a new ligand. The latter showed the distinctive characteristic to be hemilabile, leading the way of new reactivities. His coordination to palladium, followed by his deprotonation to give rise to the non-innocent nature of the complex is presented, as well as the application of the resulting complex in the context of a cycloisomerisation. Those results are reflecting the importance of the indenyl and indenediide pincer ligands besides the metal-ligand cooperativity in catalysis.

Die vorliegende Studie präsentiert eine Analyse der Integration der AhR und Wnt/beta-catenin Signalwege in den CYP1A1 Promotor sowie den regulatorischen Einfluss der Promotorlogik auf die Genexpression. Experimentell wurde diese Analyse mithilfe 29 mutagener Reporterkonstrukte des humanen CYP1A1 Promotors durchgeführt. Ein mathematisches Modell, welches eine Repräsentation des Crosstalks der Signaltransduktionswege mit einer statistisch mechanischen Beschreibung der kombinatorischen Promotorbelegung kombiniert, komplementierte den experimentellen Ansatz. Unter zusätzlicher Zuhilfenahme von gut kontrollierbaren synthetischen Promotorkonstrukten fand ich heraus, dass nur jenes Dioxin-responsive Element das sich am nächsten am Transkriptionsstartpunkt befindet, die Promotorbelegung an die RNA Polymerase kommuniziert. Außerdem beobachtete ich, dass Transkriptionsfaktoren alleine mit Transkriptionsfaktoren interagieren die mit benachbarten Bindestellen assoziieren, d.h. Interaktionen überbrücken keine größeren Entfernungen. Der Modellierungsansatz ermöglichte zudem die erfolgreiche Vorhersage einer UND-Gatter-ähnlichen Integration der beiden Signalwege in den Promotor. Für die genomische Architektur des CYP1A1 Promotors konnte ich die Signifikanz der Zielbindestelle des Wnt/beta-catenin Signalwegs innerhalb des cis-regulatorischen Region demonstrieren. Mithilfe des Modells fand ich heraus, dass diese Bindestelle am stärksten und vielfältigsten mit den restlichen Transkriptionsfaktoren interagiert. Zusätzliche konnte, im Vergleich zu dem alles-oder-nichts UND-Gatter der synthetischen Konstrukte, eine sehr viel graduellere Antwort auf die Integration der beiden Signalwege aufgezeigt werden. Abschließend wurde das physiologisch zu beobachtende Expressionsmuster von dem Modell vorhergesagt und experimentell validiert. / The study at hand presents an analysis of the integration of the AhR and the Wnt/beta-catenin signaling pathways into the CYP1A1 promoter as well as the regulatory influence of the promoter logic on gene expression. Experimentally, this analysis was conducted with the help of 29 mutant constructs of the human CYP1A1 promoter. I complemented this experimental approach with a set of mathematical models that combined a representation of the signaling crosstalk with a statistical mechanics description of the combinatorial promoter occupancy. With the help of well controllable synthetic promoter constructs I found that only the dioxin responsive element closest to the transcription start site communicates the promoter occupancy to the RNA polymerase. Furthermore, transcription factors only interact with transcription factors that associate with nearby binding sites, i.e., no long-distance binding was observed. The modeling approach subsequently enabled the successful prediction of an AND-gate-like integration of the two signaling pathways into the promoter. For the genomic architecture of the CYP1A1 promoter, I could demonstrate the importance of the Wnt/beta-catenin pathway target binding site within the cis-regulatory region. The model uncovered that this binding site is the strongest and most promiscuous interaction partner of the remaining transcription factors. In addition, a less switch-like response to the integration of the two signaling pathways as compared to the all-or-none AND-gate within the synthetic constructs could be demonstrated. And lastly, the physiological expression pattern in liver lobules could be successfully predicted by the model and experimentally verified.

<p>The glutathione transferases (GSTs) represent a superfamily of dimeric proteins involved in cellular detoxication by catalyzing the nucleophilic addition of the reduced glutathione (GSH) to the hydrophobic electrophiles. The present work focuses on the functional role of the conserved structures of GSTP1-1. The lock-and-key motif is a highly conserved hydrophobic interaction in the subunit interface of Pi, Mu, and Alpha class GSTs. The key residue (Tyr⁵⁰ in hGSTP1-1) of one subunit is wedged into a hydrophobic pocket of the neighboring subunit. The heterodimer GSTP1/Y50A was constructed from the fully active wild-type GSTP1-1 and the nearly inactive Y50A in order to study how an essentially inactive subunit influences the activity of the neighboring subunit. The results illuminate the vital role of the lock-and-key motif in modulating the GSH binding and the rate of catalysis. Additionally, the two active sites of the dimeric enzyme work synergistically. An observed water network, in hGSTP1-1 structures, connects the two active sites, thereby offering a mechanism for communication between the two active sites.</p><p>Cys⁴⁸ and Tyr⁵⁰ were targeted by mutations and chemical modifications for understanding how the α2 loop residues modulate GSH binding and catalysis. The replacement of Tyr⁵⁰ with different unnatural amino acids showed that the nature of the key residue side-chain influences the interaction with the lock structure and, consequently, the catalytic activity. The K_M^GSH, GSH affinity and protein stability can be modulated by fitting key residue into the lock cavity of the neighbor subunit and, consequently, restriction of the flexibility of the α2 loop. Optimization of the interaction between the key residue and the lock-cavity increases k_cat. Also, the crystal structure of the Cys-free variant was determined. The result indicated that Cys⁴⁸ restricts the flexibility of the α2 loop by interacting with surrounding residues and, consequently, contributes to GSH binding and protein stability.</p>

Die Beeinflussung optischer Eigenschaften durch Bestrahlung stellt eine Grundlage für die Herstellung anisotroper optischer Komponenten dar. In dünnen Schichten von Azobenzen-Polymeren kann optische Anisotropie durch linear polarisierte Bestrahlung induziert oder modifiziert werden. Ziel der Arbeit war es, wesentliche Struktur-Eigenschafts-Beziehungen zum Prozess der Photoorientierung zu erarbeiten, um so eine Optimierung der Materialien für verschiedene Anwendungen ermöglichen. <br><br> In isotropen Schichten flüssigkristalliner und amorpher Azobenzen-Polymeren wird das Ausmaß der induzierten optischen Anisotropie günstig durch eine Donor/Akzeptor-Substitution in 4,4'-Position beeinflusst. Die Induktionsgeschwindigkeit ist in Schichten flüssigkristalliner Polymeren deutlich geringer, jedoch lassen sich höhere Werte der Doppelbrechung und des Dichroismus erreichen. In Copolymeren bewirkt die Photoorientierung der Azobenzen-Seitengruppen eine kooperative Orientierung von formanisotropen Seitengruppen. Die Mesogenität der nicht-photochromen Seitengruppen erhöht das Ausmaß der induzierten optischen Anisotropie. Die Stabilität der induzierten Doppelbrechung und des Dichroismus wird durch diese Gruppen gesteigert. <br><br> In Schichten flüssigkristalliner Polymeren wird die induzierte optische Anisotropie beim Tempern im Bereich der Mesophasen erheblich verstärkt. Dabei reicht ein geringes Maß an induzierter Anisotropie aus, um Doppelbrechungs- und Dichroismuswerte zu erzielen, wie sie für LC-Domänen typisch sind. <br><br> In orientierten Schichten von Azobenzen-Polymeren wird das Resultat der linear polarisierten Bestrahlung durch die Stärke der anisotropen Wechselwirkungen in den flüssigkristallinen Domänen oder den LB-Multilayern bestimmt. Eine lichtinduzierte Reorientierung kann nur erreicht werden, wenn diese Wechselwirkungen überwunden werden können. Erfolgt eine Photoreorientierung in den orientierten Polymerschichten, werden in Copolymeren formanisotrope Seitengruppen ebenfalls kooperativ reorientiert. Eine vorgelagerte UV-Bestrahlung kann durch Erzeugung eines hohen Anteils an nicht-mesogenen Z-Isomeren die anisotropen Wechselwirkungen stark schwächen und so die Seitengruppen entkoppeln. Aus diesem Zustand erfolgt die Photoreorientierung mit einer der Photoorientierung in isotropen Schichten vergleichbaren Effizienz. <br><br> Die erarbeiteten Struktur-Eigenschafts-Beziehungen liefern einen Beitrag zur Optimierung derartiger Materialien für Anwendungen in den Bereichen optischer Funktionsschichten, holographischer Datenspeicherung oder der Generierung von Oberflächenreliefgittern. / The modification of optical characteristics by irradiation represents a basis for the creation of anisotropic optical components. In thin films of azobenzene polymers optical anisotropy can be induced or modified by linearly polarized irradiation. The goal of the work was it to compile substantial structure property relations of the photoorientation process in order to support an optimization of the materials for different applications. <br><br> In isotropic films of liquid crystalline and amorphous azobenzene polymers the extent of the induced optical anisotropy is increased by a donor/acceptor substitution in 4,4'-position. The induction speed is clearly smaller in films of liquid crystalline polymers, however higher values of birefringence and dichroism can be reached. In copolymers the photoorientation of the azobenzene side groups causes a cooperative orientation of form-anisotropic side groups. The mesogenity of the non-photochromic side groups increases the extent of the induced optical anisotropy. <br><br> The stability of the induced birefringence and dichroism is increased by these groups considerably. In films of liquid crystalline polymers the induced optical anisotropy is substantially amplified on annealing within the range of the mesophases. In this way, a small ratio of induced anisotropy is sufficient, in order to obtain birefringence and dichroism values as typical for LC domains. <br><br> In oriented films of azobenzene polymers the result of the linear polarized irradiation is determined by the strength of the anisotropic interactions in the liquid crystalline domains or the LB multilayers. A light-induced reorientation can be only achieved, if these interactions can be overcome. If a photoreorientation takes place in the oriented layers of copolymers, form-anisotropic side groups are cooperatively reoriented. An initial UV irradiation can strongly weaken the anisotropic interactions by generating of a high fraction of non-mesogenic z-isomers and decouples in this way the side groups. From this state the photoreorientation proceeds with an efficiency one comparable to the photoorientation in isotropic films. <br><br> The compiled structure property relations supplies a contribution for the optimization of such materials for applications in the fields of optical function films, holographic data storage or the generation of surface relief gratings.