23 search hits
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The Aggregation Behavior of Mixture of Alkylmethylaminoxides with Their Protonated Analogues in Aqueous Solution
(2005)
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Yuji Yamashita
- The C12C8MAO aqueous solution at 100 mM consisted of two phases which were optically isotropic and low viscous. Addition of chloric acid induced a phase transition, and the following lamellar (L-alpha) phase was formed in the range of low protonation degree, X = 0.007 – 0.35. When the surfactant was protonated further (X > 0.35), the single L-alpha phase separated again into two isotropic phases. The abnormal phase sequence could be interpreted by the result that mixtures of protonated and non-protonated C12C8MAO were more surface active than each component. The surface and interfacial measurements showed synergism of mixing two components. This synergistic effect arised from the peculiar interaction of hydrogen bonding between protonated and non-protonated head groups. This short-range interaction would cause the C12C8MAO molecules to be more lipophilic with protonation, resulting in the phase separation at high protonation degree. The SAXS measurements in the L-alpha phase also showed the synergistic effect between the head groups. The rheological measurements and microscope observations demonstrated that the morphologies of L-alpha phase could be controlled by preparation routes. It was found that the vesicles were transformed into the classical lamellar phase by the simple process of heating and cooling through the phase transition (L-alpha <-> L-alpha;/L1) temperature. Furthermore, the classical, planar lamellar morphology could be prepared by means of kinetic protonation of the C12C8MAO molecules using hydrolysis reaction. Any classical L phase was modified to the vesicle form under shearing, and its transformation was irreversible in terms of shear force. Various acids were treated as protonation agent in 100mM C14DMAO aqueous solution, and their contributions to the viscosity of solution were examined ranging X = 0 to 1. It was elucidated that the aggregate structure remarkably depended on the ion-pair (counte-ion) thermodynamics. For the Cl-, HCOO-, and H2PO4- ions, no remarkable structural change took place with increasing protonation. For the Br-, NO3-, Oxalate, Tartarate, Tartronate, and SO42- ions, the small micelles of C14DMAO grew up with protonation. For the ClO4-, SCN-, and Salicylate ions, the L-alpha phase was formed with protonation. The interfacial curvature thus followed mostly the sequence of Hofmeister series, while the sulfuric ion was completely excluded from the series because of its divalency. The viscosity change was interpreted quantitatively by the hydration free energy (Delta-Ghyd) of counter-ion. However the divalent counter-ions did not arrange in the same order as the monovalent ones, that could be considered to arise from (1) depressing pKa of C14DMAO with increasing amount of acid and (2) electrostatic force within the diffuse double layer. For different counter-ions, the characteristic scaling laws of viscosity evolution against X were observed. The scaling laws also obeyed Delta-Ghyd. Sulfate, however, could not be manipulated by Delta-Ghyd: regardless of its strong hydrophilicity (high Delta-Ghyd), the excess amount of SO42- caused the micelles to grow up. The micelle growth therefore would be attributed by counter-ion condensation onto the micelles rather than hydrophilicity itself. Comprehensively it was proved that Delta-Ghyd and the ionic valency (electrostatic force) of counter-ion played strikingly significant roles in the structural properties. Trifluoro acetic acid CF3COOH behaved as hydrophobic acid, however, the viscosity trend could stand in neither the sequence of Delta-Ghyd nor the Hofmeister series. The interfacial tension measurments suggested that the CF3COO- ion was incorporated into the micelle, behaving like co-surfactant. Synergism on mixing was also observed in the CF3COOH system, by which the minimal CMC was obtained. The interaction parameter beta indicated that CF3COOH caused the stronger synergistic effect than HNO3. However, some experiments showed the successively hydrophobic C14DMAO with protonation. This would be due to the orientation of hydrophobic trifluoro group in the palisade layer of micelle. The increasing volume fraction of hydrophobic moiety in the surfactant molecule (OleylDMAO) built up the bilayer structure even using the strongly hydrated acids. The L-alpha formation was referred to synergism because the isotropic micellar phase was present on both the sides of high and low protonation degrees. The L-alpha phase melted on elevating temperature, and the subsequent L1 phase was highly viscoelastic. The L-alpha - L1 transition temperatures for different acids were almost correlated by the enthalpy of hydration (DeltaHhyd). And the viscoelastic properties of the L-alpha phases were dependent on the Hofmeister series.
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Synthese, Charakterisierung und Reaktivität von Platin(II)-Komplexen mit Cycloheptatrienylphosphan-Liganden
(2005)
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Bettina Ullmann
- Tri(1-cyclohepta-2,4,6-trienyl)phosphan P(C7H7)3 (1) kann als ein mehrzähniger Ligand fungieren, da es sowohl über das Phosphoratom als auch über die mittlere Doppelbindung der Siebenringe an ein Zentralmetall koordinieren kann. Dabei war in Komplexen, bei denen das Phosphan zweizähnig gebunden ist, dynamisches Verhalten bezüglich der Koordination der Doppelbindung zu beobachten. Deshalb erschienen die NMR-Signale gemittelt und zum Teil sehr breit. Zusätzlich wurde die Reaktivität dieser Komplexe durch die dynamischen Prozesse eingeschränkt. Daher sollten im Rahmen dieser Arbeit neue Cycloheptatrienylphosphane synthetisiert werden, die zweizähnig an das Platin gebunden sind und geringe Dynamik zeigen. Ein Vertreter solcher Phosphane ist das Cycloheptatrienyl(diphenyl)phosphan Ph2P(C7H7) (3), welches über einen optimierten Syntheseweg hergestellt werden konnte. Bei der Darstellung zwei neuer Phosphane, dem MeP(C7H7)2 (6) und dem Me2P(C7H7) (8), wurde genutzt, dass Cycloheptatrienylphosphoniumsalze durch eine reduktive Eliminierung von Tropyliumbromid mit anschließender Umsetzung mit LiAlH4 stets wieder in ein Phosphan zurückgeführt werden können. Deshalb wurde 1 mit Methyliodid versetzt und anschließend ein C7H7-Ring eliminiert. Durch nochmaliges Umsetzen von 6 mit Methyliodid und anschließender Reduktion erhält man das Dimethyl(cycloheptatrienyl)phosphan (8), das nur einen Siebenring enthält und dadurch fest an das Platin koordiniert. MeP(C7H7)2 (6) ist ein anschauliches Beispiel für 1H- und 13C-NMR-Spektren von diastereotopen =CH-Einheiten des Siebenringes. Neben der Verwendung der Phosphane als Liganden wurden sie mit den elementaren Chalkogenen, O, S, Se, zu Chalkogeniden umgesetzt. Um schließlich die Koordinationschemie der drei dargestellten Phosphane 3, 6 und 8 zu untersuchen, wurde (cod)PtCl2 mit dem entsprechenden Phosphan versetzt (12, 14, 15). Dadurch wird cod durch einen Cycloheptatrienylphosphanliganden substituiert. Die Phosphane koordinieren über das Phosphoratom und über die mittlere Doppelbindung des Siebenringes an das Metall. Wie erwartet zeigen die C7H7-Ringe von 3 und 8 keine dynamischen Prozesse in ihren Platinkomplexen. Die Molekülstruktur für 12 wurde mittels Röntgenstrukturanalyse bestimmt. Dagegen zeigt 14 in den 1H- und 13C-NMR-Spektren wie [(C7H7)2P(C7H7)]PtCl2 (2) nur gemittelte Signale. Wird das Phosphan 3 im zweifachen Überschuss zum (cod)PtCl2 gegeben, wird der Diphosphankomplex mit zwei einzähnig koordinierten Phosphan-Liganden erhalten, welche cis-ständig am Platin gebunden sind. Wie bereits für 2 beschrieben, ist es möglich, den zum Phosphor-Atom cis-ständigen Chloro-Liganden durch eine Alkinylgruppe zu substituieren. Die Umsetzung von 2 mit Me2Sn(CCR)2 bei Raumtemperatur führt interessanterweise zu Di(alkin-1-yl)platin-Komplexen. Vorher mussten diese über eine Zwischenstufe, dem (cod)Pt(CCR)2, dargestellt werden. Der kristalline Komplex 22 wurde mittels Röntgenstrukturanalyse charakterisiert. Für die Reaktionen mit MeLi wurde die Synthese der Komplexe [(C7H7)2P(C7H7)]PtCl(CCR) wiederholt. Dabei konnte für 27 (R = SiMe3) ein Einkristall erhalten werden, der für eine Röntgenkristallstrukturanalyse geeignet war. Der Chloro-Ligand sollte leicht über Substitutionsreaktionen ersetzt werden können. Allerdings wurden mit MeLi nur Produktgemische erhalten, und diese Produkte wurden mithilfe von verschiedenen NMR-Versuchen charakterisiert. Die Produktverteilung zeigt an, dass die Reaktion unter nucleophiler Addition des Methylanions und anschließender Eliminierung von LiCl, MeLi oder LiCCR abläuft, wobei das Letztgenannte dann auch an der Reaktion teilnimmt. Bei der Umsetzung von 27 mit einem Überschuss an MeLi wird ein weiteres Produkt (44) erhalten, welches an der CC-Gruppe methyliert wurde und durch Röntgenstrukturanalyse charakterisiert werden konnte. Die 1,1-Organoborierungsreaktionen der Alkin-1-yl(chloro)platin-Komplexe laufen sehr langsam bis gar nicht ab. Ein möglicher Grund dafür ist, dass der Alkinyl-Rest trans zur Olefin-Einheit steht. Sie übt im Vergleich zum Phosphor-Atom offenbar einen geringeren trans-Einfluss aus und die Pt-C-Bindung ist für die Organoborierungsreaktion nicht hinreichend polar. Aufgrund des höheren trans-Einflusses des Phosphor-Atoms reagieren die Di(alkin-1-yl)platin-Komplexe an der zum Phosphor trans-ständigen Alkinyl-Gruppe mit BEt3. Dabei entstehen mehrere neue Verbindungen, von denen bisher nur wenige charakterisiert werden konnten. Ein solches Produkt ist 21A, für das zahlreiche NMR-Daten vorliegen. Wenn der Alkinyl-Rest mit einer Trimethylsilyl-Gruppe substituiert ist, kommt es zur Bildung zweikerniger Komplexe (22D, 43D), welche interessante NMR-Spektren liefern. Die spektroskopischen Untersuchungen der Umsetzung von 2 mit LiSnMe3 liefern für ein Produkt sehr komplizierte NMR-Spektren, da sich hier vermutlich ein Platin(II)-Komplex (46) mit vier Stannyl- und einem Phosphan-Liganden bildet.
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Neue Alkin-1-ylsilane und -germane, Sila- und Germacyclopentadiene – Charakterisierung mittels Multikern-NMR-Spektroskopie
(2005)
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Stefan Bayer
- 1-Metalla-2,4-cyclopentadiene (Metallole) wie z. B. Silole und Germole sind seit 1959 bekannt. Ihre Darstellung war allerdings nur durch vielstufige Synthesen zu realisieren und auf C(2–5)-Aryl-substituierte Derivate begrenzt. Es gibt bisher kaum Methoden, die in hoher Ausbeute zu verschiedenartig substituierten Silolen und Germolen führen. Die 1,1-Organoborierung von Di- und Poly(alkin-1-yl)metall-Verbindungen bietet hier eine vielseitig einsetzbare Alternative, nachdem bereits Stannole und Plumbole auf diesem Weg hergestellt wurden. Die Bezeichnung 1,1-Organoborierung erklärt sich damit, dass sowohl R2B als auch R am selben Kohlenstoff (alpha) enden. Für diesen Reaktionstyp wurden zahlreiche Alkin-1-ylsilane und -germane darge-stellt und anhand der Multikern-NMR-Spektroskopie untersucht. Besonders gut lässt sich der Verlauf der Substitution durch 29Si-NMR-Spektroskopie detektieren, selbst in verdünnten Lösungen und komplexen Mischungen. Bei der Darstellung wurden normalerweise Produktmischungen erhalten und destillativ aufgearbeitet. „Gleich-substituierte“ Dialkin-1-yl-Verbindungen mit zwei oder mehr gleichen Resten konnten so erhalten werden. „Gemischt-substituierte“ Alkin-1-yl-Verbindungen mit zwei oder mehr verschiedenen Alkin-1-ylfragmenten konnten durch sukzessive Alkinylierung von Silicium-Chloro-Verbindungen dargestellt werden. Monoalkin-1-yl-Verbindungen bilden borylierten E-Alkene. Setzt man Dialkin-1-ylsilane ein, so kann das E-Alken mit der verbleibenden Alkin-1-yl-Funktion eine weitere 1,1-Organoborierungsreaktion eingehen. Der intramolekulare Charakter des zweiten Reaktionsschrittes, verbunden mit den extremen Reaktionsbedingungen, führt in einer schnellen und selektiven 1,1-Vinylborierung unter irreversiblem Ringschluss zu den Silolen. Analog hierzu reagieren Alkin-1-ylgermane mit BEt3 im Allgemeinen bereits bei niedrigeren Temperaturen zu Germolen, die mit anderen Verbindungen verunreinigt sind. Da viele Derivate der Silicium- und Germaniumchemie über Halogen- und Wasserstoff-Intermediate synthetisiert werden, strebt man Silole und Germole an, die eine oder beide dieser funktionellen Gruppen am Metall tragen. Die Organoborierung toleriert während der Reaktion H- und Cl-Substituenten direkt am Silicium. Durch Überwachung der Reaktion mittels 1H-, 11B-, 13C- und 29Si-NMR-Spektroskopie konnten die Reaktionsbedingungen für inter- und intramolekulare 1,1-Organoborie-rung erfasst und Zwischenstufen vor der Bildung der Silole detektiert werden. Unter gleichen Reaktionsbedingungen konnten Dichloro- und Dihydrido-Silolderivate dargestellt werden. In allen Fällen waren die Endprodukte Silole, die als luft- und feuchtigkeitsempfindliche, ölige Flüssigkeiten isoliert wurden. Die Verfügbarkeit von „ungleichmäßig-substituierten“ Alkin-1-ylsilanen ermöglichte die Darstellung „ungleichmäßig-substituierter“ Silole. Erstmals war es möglich, diese besonderen Substitutionsmuster in Position 1–5 zu erhalten. Besonders die funktionellen Gruppen am Silicium- und Germanium-Atom versprechen weitere Umsetzungen. Enthält das Edukt mehr als zwei Alkin-1-yl-Gruppierungen, so muss die 1,1-Organo-borierung nicht mit der Ausbildung des ersten Ringsystems enden. Unter diesen schroffen Reaktionsbedingungen reagieren verbleibende Alkin-1-yl-Funktionen mit BEt3 unter zusätzlicher Organoborierung. Dies führt zu verschiedenen Isomeren (E / Z) mit und ohne Silicium-Wasserstoff-Bor-Brückenbindung. Verglichen mit dem anderen Isomer zeigt das Brücken-H-Atom stark unterschiedliche Si-H-Kopplungskonstanten, Si-H-Valenzschwingungen (IR) und 1H-NMR-Werte. Die 1,1-Organoborierung von Tetraalkin-1-yl-silanen und -germanen mit BEt3 ergab durch zwei nacheinander erfolgende Ringschlüsse 1,1’-Spiro-di-silole oder entsprechende -germole. Unter den gewählten Reaktionsbedingungen gelingt die Umsetzung von erhaltenen tBu-Derivaten auch bei sehr langen Reaktionszeiten nur teilweise. Die Chloro-Funktion ist eine attraktive Abgangsgruppe bei Substitutions-Reaktionen. Auf diese Weise konnten verschiedene Organyl-Reste direkt am Siliciumatom eingeführt werden. Eine große Anzahl an [4+2]-Cycloadditionen von C-phenylierten Silolen mit Dieno-philen, die zur Bildung von 7-Sila-norbornenen oder 7-Sila-norbornadienen führen, wurden bereits beschrieben. Mit DMADC oder TCE und wurden schnell und unter relativ milden Reaktionsbedingungen analoge Diels-Alder-Produkte dargestellt. Mit H und Cl am Siliciumatom können [4+2]-Cycloadditionen zu zwei verschiedenen Isomeren führen. Die Diels-Alder-Reaktion dieser Silole mit DMADC und TCE führte entweder selektiv zum anti-Isomer, oder man erhielt, nicht vorhersehbar, Gemische aus beiden Isomeren. NOE-Messungen bestätigten diese Konfiguration, und die Struktur wird durch DFT-Berechnungen unterstützt. Die neu erhaltenen anti-Isomere zeigten sich bei Berechnungen um ca. 16 kJ/mol thermodynamisch stabiler. Errechnete 29Si-Verschiebungen ergaben zu experimentellen Daten analoge Trends.
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Structural Analysis of Cylindrical Particles by Small Angle X-ray Scattering
(2005)
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Li Li
- The objective of this work is to analyze nano-scaled cylindrical particles by small angle X-ray scattering (SAXS). Three systems with cylinder-shaped particles: (1) Laponite particles in aqueous solutions, (2) Poly(carbon suboxide) particles in binary water/DMF solutions, and (3) Suprastructural aggregates of coil-ring-coil block copolymers in cyclohexane, have been studied by SAXS performing either a Kratky-Compact-Camera in our laboratory or ID2 beamline of the European Synchrotron Radiation Facility (ESRF) in Grenoble. The synthetic clay particles, Laponite RD, have been chosen as ideal disc-shaped model particles. In combination of SAXS with static light scattering, the scattering intensities of a concentration serial (volume fraction from 0.0002 to 0.0016) were measured in almost three orders of magnitude of the scattering vector q. Through extrapolation of concentration the scattering intensity at vanishing concentration, i.e. the form factor P(q) of particles was achieved. It shows q-2 decay at intermediate q range, which indicates that the shape of single Laponite particle in aqueous solution is platelet. The plateau of the form factor at low q range implies that there is no aggregate or cluster structure, and the Laponite particles are dispersed completely under the investigated conditions. More detailed structural information was then obtained by fitting of P(q) with disc model. The radii of the discs exhibit a large polydispersity. A radius of 10.5 nm with Schulz-Zimm distribution of Rw/Rn = 1.5 (where Rw and Rn denote weight and number average radius, respectively) was found to fit the form factor perfectly. The thickness of one single platelet was determined to be 0.9 nm. The weight averaged molecular weight and radius of gyration were determined to be 930 kg/mol and 13.4 nm, respectively. The inter-particle interactions of Laponite particles were investigated by the structure factor S(q), from which the effective diameter of interparticle interactions deff was determined for the first time. The strong electrostatic Coulomb repulsion between charged Laponite particles was attributed to the much higher value of deff (= 46 nm), in comparison to 2Rg (= 27 nm). The recently developed multicomponent interaction site model was performed by Harnau to predict these experimental structure factors. An effective potential of interaction, which pays attention to a screened Coulomb interaction as well as an attractive interaction, leads to the best description of the model to the experimental data. By means of SAXS, the size of synthetic polymer carbon suboxide ((C3O2)n) dissolved in binary water/DMF solutions was determined for the first time with radius of gyration Rg = 1.7 nm and molecular weight Mw = 2760 g/mol, which corresponds to a polymerization’s degree of about 40. This value is much larger than literature one (5-10). The form factor of polymer carbon suboxide can be described by a semiflexible chain model. The radius of gyration in cross-section RC and molecular weight per unit length ML were obtained to be 0.3 nm and 350 g/(mol.nm), respectively, which can confirm the fact that the chemical structure of poly(carbon suboxide) is repeated pyronic ring, as suggested in most literatures. Thus the structure and size of polymer carbon suboxide were characterized completely by SAXS. Finally, SAXS was employed to analyze a suprastructural aggregation system derived by self-assembly of coil-ring-coil block copolymers. This is a newly synthesized subclass of rod-coil block copolymers composed of a nanometer-sized shape-persistent macrocycle and two covalently attached polystyrene (PS) coils. The solubility of the rigid ring is largely enhanced due to the attachment of the flexible side groups. With suitable length of the flexible side groups (Mw (PS) = 2500 g/mol) the block copolymers can form colloidal-sized aggregates in selected solvent cyclohexane, which were concluded to be of cylindrical shape with the rigid rings packing densely in a tubular way and the flexible side groups arranging outside of the ring. Such aggregated cylinder brushes can be further confirmed to exist as a mixture of cylinder bundles by analyzing the local structural parameter ML (= 25730 g/(mol.nm), molecular weight per nm length of cylindrical objects). In comparison of this value with M0 (= 6500 g/mol, molecular weight of single coil-ring-coil block copolymer) and d (= 0.6 nm, distance of adjacent densely packed rings), the number fraction of coexisted single cylinder, bi- and tri-cylinder bundles was resulted to be 1:1:2. Through fitting by using approximated circular cylinder model the radius of single cylinders was determined to be 2.6 nm (polydispersity 20 percent) with a hollow inside of radius of 1.2 nm.
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Mechanismen molekularer Erkennung: Charakterisierung der molekularen Details der Wechselwirkungen des herpesviralen Tip-Proteins mit SH3-Domänen
(2005)
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Finn Bauer
- Die Wechselwirkungen des Tip-Proteins aus Herpesvirus saimiri mit der T-Zell-spezifischen Kinase Lck sind entscheidend für die Aktivierung der Kinaseaktivität und nehmen somit eine zentrale Rolle bei der T-Zell-Transformation ein. Bis heute war es weder durch Röntgenkristallografie, noch durch NMR-Spektroskopie möglich, eine hochaufgelöste Struktur des LckSH3-Tip-Komplexes zu erhalten. Eine Möglichkeit dieses Probleme zu lösen, ist die Verwendung von stärker bindenden Interaktionspartnern, die sich oft besser für strukturelle Studien eignen. Aus diesem Grund wurde in der vorliegenden Arbeit sowohl der Komplex von Tip mit LynSH3 charakterisiert, als auch eine Punktmutante (P17G) der LckSH3 entworfen, die eine höhere Affinität zu Tip aufweist. Die Strukturinformationen, die aus der Charakterisierung dieser Systeme erhalten werden konnten, sollten als Basis für eine zuverlässige Modellierung des LckSH3-Tip-Komplexes dienen. Analysen ergaben, dass die im Vergleich zur LckSH3 erhöhten Ligandenaffinitäten von LynSH3 und LckSH3_P17G auf unterschiedlichen Mechanismen basiert. Die im Vergleich zur wildtyp LckSH3 um nahezu eine Zehnerpotenz stärkere Bindung von LckSH3_P17G konnte durch schnellen Mischmethoden zumindest teilweise auf eine schnellere Komplexbildung zurückgeführt werden. Eine detaillierte Erklärung dieses Unterschieds auf struktureller Ebene konnte durch Moleküldynamiksimulationen des Wildtyps und der Mutante der LckSH3 erhalten werden. So korreliert die schnellere Komplexbildung der Mutante gut mit einer stärkeren Population der 'bindungsaktiven' Konformation während des Simulationszeitraums. Dabei zeichnet sich LckSH3_P17G nicht nur durch eine erhöhte konformationelle Abtastrate der 'bindungsaktiven' Konformation aufgrund gesteigerter RT-Schleifenflexibilität aus, sondern zeigt eine Zunahme der Gesamtflexibilität der SH3-Domäne im ps-s-Bereich auf. Weiterhin konnten durch die Analyse der thermischen Entfaltung mittels CD-Spektroskopie und MD-Simulationen eine globale Destabilisierung und ein veränderter Entfaltungsweg für die P17G-Mutante der LckSH3 gezeigt werden. Der LynSH3-Tip-Komplex konnte in hoher Auflösung mittels NMR-Spektroskopie bestimmt werden. Analysen der Komplexstruktur ergaben, dass nicht nur die Prolinhelix, sondern auch zusätzliche COOH-terminal flankierende Reste des Liganden an der Bindung beteiligt sind. Eine besondere Rolle nimmt dabei L186 ein. Es bindet in eine hydrophobe Tasche aus H41, W44 und F57 auf der Oberfläche der LynSH3. Ausgehend von der LynSH3-Tip-Komplexstruktur konnte unter der Einbeziehung NMR-spektroskopischer Daten der LckSH3-Tip-Wechselwirkungen ein molekulares Modell des LckSH3-Tip-Komplexes erstellt werden. Obwohl die Bindung von L186 in die hydrophobe Tasche auf der Oberfläche für beide SH3-Domänen experimentell nachgewiesen werden konnte, zeigten Fluoreszenztitrationsexperimente mit einem verkürzten Tip-Peptid, dass L186 aus Tip nur im Komplex mit LynSH3 einen signifikanten Einfluss auf die Affinität hat. Weitere Untersuchungen ergaben, dass L186 bei Bindung in die hydrophobe Tasche der LynSH3 nahezu vollständig von H41 bedeckt wird. Das Serin an strukturell ähnlicher Position in LckSH3 bedeckt hingegen nur einen kleinen Teil von L186. Die zusätzlichen Wechselwirkungen senken die Geschwindigkeitskonstante der Dissoziation des Komplexes, was in guter Übereinstimmung mit dem unterschiedlichen Austauschverhalten auf der NMR-Zeitskala steht. Zusammenfassend konnte in dieser Arbeit neben der Aufklärung der molekularen Grundlagen der Wechselwirkungen der LckSH3 mit dem herpesviralen Tip-Protein gezeigt werden, dass Affinität auch bei sehr nahe verwandten Proteinen auf unterschiedliche Weise erzielt werden kann. Weiterhin unterstreichen die Ergebnisse dieser Arbeit die Notwendigkeit der gleichzeitigen Charakterisierung der Struktur und Dynamik makromolekularer Interaktionen. Erst dieses kann zu einem detaillierteren Verständnis der Mechanismen molekularer Erkennung und darauf aufbauend zukünftig zu verlässlicheren Vorhersagen von molekularen Interaktionen führen.
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Synthesis and Characterization of Cationic Spherical Polyelectrolyte Brushes
(2005)
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Yu Mei
- In this thesis, the preparation of cationic spherical polyelectrolyte brushes by photo-emulsion polymerization has been presented for the first time. The structure and properties of the cationic SPB, have been comprehensively investigated by dynamic light scattering and atomic force microscopy, and compared with anionic SPB. Moreover, both of them were tested as dual retention-aid in conjunction with cationically modified polyacrylamide in papermaking. The interaction of SPB with negatively charged surfaces was investigated by AFM operated in Tapping Mode. It was demonstrated that the negative SPB forms two-dimensional aggregates of densely packed polymer particles, which can be explained by a particle-particle interaction dominating the repulsive interaction of the particles with the mica substrate. The positively charged SPB exhibits a completely different particle-surface interaction behaviour from that of negatively charged SPB. Here network-like structure films of dried particles without long-range 2D order are formed, which is due to the strong attractive particle-surface interaction of positively charged polyelectrolyte chains in the shell of the cationic SPB particles. These chains spread over the negatively charged mica surface and anchor the particles. The swelling behaviors of cationic and anionic SPB as a function of ionic strength in the system were studied by DLS. Adding more and more salt leads to a strong shrinkage of the surface layer as expected for polyelectrolyte brushes. For some ions, however, high salt concentrations may lead to a re-swelling of the brush layer in case of the cationic systems. This points to specific interactions of the counterions with the PATAC chains. This strong specific interaction between the counterions and the attached polyelectrolyte may even lead to flocculation of the particles at intermediate salt concentration. Surprisingly, for sodium iodide and magnesium sulfate the solubility increases again if the salt concentration is raised to 1 mol/l. At lowest ionic strength electrostatic interaction prevails and the brush layer is swollen in all cases by the osmotic pressure of the counterions. Intermediate salt concentrations lead to a partial screening of the electrostatic interaction and to a shrinkage of the brush layer. This effect can be well captured by the theory of Hariharan et al.. In case of cationic brushes, however, the shrinkage becomes very pronounced around salt concentrations of 0.1 mol/l. In some cases there is even a collapse of the surface layer due to specific interactions between the polyion and the counterions. Cationic systems re-swell if immersed in concentrated salt solutions. This is observed for monovalent as well as for divalent counterions. The analysis of the reduced excluded-volume parameter v/(lKl2) suggests that there is an adsorption of the counterions at high salt concentrations. The salting-in behavior thus finds an explanation in the increase of v due to the adsorption of salt ions. All data demonstrate that specific effects of different counterions lead to a behavior of the brush layer not expected from a purely electrostatic model. All specific effects seen at high concentrations of added salt can be explained by the increase of the reduced excluded-volume parameter which is due to the adsorption of salt ions. Cationic and anionic SPB were tested as dual-component retention system in combination with cationically modified polyacrylamide, and compared with the traditional "microparticle" system in which bentonite acts as secondary flocculant. The anionic SPB-based system shows high flocculation efficiency as tested under sheared conditions using a dynamic drainage jar. The high retention level of the SPB dual-component retention system can be explained by the enlarged specific surface area and cation exchange capacity of SPB, which are caused by grafted flexible polyelectrolyte chains. Furthermore, chemically grafted polyelectrolyte brushes show more advantages than bentonite, which may further delaminate upon dilution and cause CaCO3 fillers partially detach from the fiber. Images from field emission scanning electron microscopy of difference stages of retention testes suggest that the flocculation mechanism of anionic SPB and CPAM as dual-component retention system can be summarized in three steps: at first, excessive CPAM were added to bind cellulose fibers and CaCO3 fillers and formed loose macroflocs with positive charges; then the agglomerations were broken into microflocs by strong shear strength; at last, anionic SPB with negative charges were added and caused much finer and denser flocs, thus creating paper sheets with better optical appearance due to higher homogeneity. Images from FESEM images and AFM support the model of anionic SPB’s acting as a particle bridge between fibers and CaCO3 fillers.
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Synthesis and characterization of bifunctional materials for electro-optical applications
(2005)
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Katja Peter
- This thesis is dealing basically with synthesis of organic materials for electro-optical applications and interfacial modifications in such applications. It is structured into two major sections each of which presenting a new concept of bifunctional materials: I. Bifunctional Ru(II) dyes carrying hole transport units II. Fully functionalized AB-diblock copolymers I. Bifunctional Ru(II) dyes carrying hole transport units for interface modification in solid-state dye-sensitized nc-Titaniumdioxide solar cells The main concern in this part of this work was the improvement of the Titaniumdioxide/ dye / hole conductor interfaces in solid-state dye-sensitized Titaniumdioxide solar cells. Therefore novel concepts of bifunctional materials carrying light absorbing Ru(II) dye centre as well as hole transport triphenylamine moieties were developed for the following reasons: • Improvement of wetting between polar Ru(II) dye layer and the non-polar spiro-hole conductor. • Spatial separation of excited dye cation centre (HOMO) away from the Titaniumdioxide surface. • Retardation of recombination of holes in the dye molecule with electrons in Titaniumdioxide. In order to reach these goals, novel bipyridine ligands carrying hole transport units (bpy-TPA 6 and bpy-TPD 11) with bipyridine centre for coordination of a transition metal atom have been prepared. In a metallation step a Ru(II) dye was created which carries holetransport moieties as antenna groups. The new, bifunctional materials have been characterized and their potential for application in electro-optical devices have been investigated. II. Fully functionalized AB-diblock copolymers carrying hole transport and NLO-dye blocks Another synthetic approach of this thesis was to explore the suitability of the polymerization technique ATRP for the synthesis of fully functionalized AB-diblock copolymers for photorefractive applications. For this purpose, 4-bromostyrene was polymerized via ATRP using alpha-chloromethylpropionate as initiator and CuCl / PMDETA as catalytic system yielding poly(4-bromostyrene) macroinitiators (19 a – e). The control of this polymerization reaction was verified by GC analysis before the macroinitiators had been used to initiate the polymerization of HEMA-TBDMS which was applied as monomer for the second block. By this method, AB-diblock copolymers have been prepared consisting of a poly(4-bromostyrene) block and a poly(HEMA-TBDMS) block (21). Polymeranalogous reaction yielded the desired fully functionalized block copolymers carrying hole transport and NLO-dye block.
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Generation of metal nanoparticles in spherical polyelectrolyte brushes and their application in heterogeneous catalysis
(2005)
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Geeta Sharma
- This thesis describes synthesis of spherical cationic polyelectrolyte brushes by “grafting from” technique. The spherical polyelectrolyte brushes have been used as “nanoreactors” for the synthesis of metal nanoparticles (Gold, Platinum and Silver). The catalytic activity of the resulting metal/polymer nanocomposites has been tested by heterogeneous hydrogenation of carbonyl groups. The synthesis of the cationic spherical polyelectrolyte brushes is achieved by a three-step procedure. Firstly, cationic polystyrene core particles are synthesized by emulsion polymerization by using a cationic surfactant and a cationic thermal initiator. In the second step, a thin layer of photoinitiator is generated around the particles by addition of the photoinitiator under “starved conditions” . The photoinitiator is a monomer, which is added when the formation of core particles is in the last stage. The photoinitiator polymerizes with the styrene to give a covalently bound photoinitiator. In the third and last Step, the brushes are grafted on the core particles by photoemulsion polymerization, where the initiation is triggered by UV/VIS radiation. The important parameters of the brushes- contour length and grafting density are determined by the cleavage of the chains from the surface by alkaline hydrolysis. The ester functionality within the photoinitiator is hydrolyzed under the harsh conditions of hydrolysis. The cleaved chains are analyzed to determine the molecular weight. The grafting of the charged polymeric chains stabilizes the colloids even under unfavorable conditions like high ionic strength and high pH. The behavior of brushes is investigated at different ionic strength and pH. At different ionic strengths brushes show three regimes- osmotic, salted and neutral regime. At increasing ionic strength, a shrinking in brush thickness is observed due to the screening of ionic charges. The cationic brushes (polyamino ethylmethacrylate hydrochloride and poly vinylbenzylamine hydrochloride) bear protonated amine functionality. The brushes are classified as annealed brushes as they are sensitive towards pH. The brushes lose the protons at high pH, to result in the uncharged brushes, causing them to shrink, which is followed by dynamic light scattering. Polyaminoethylmethyacrylate brushes are used as nanoreactors to synthesize the metal nanoparticles of gold and platinum. The water-soluble metal salts are to introduce metal ions in the brushes. The negatively charged metal ions (AuCl4- and PtCl6-2) interact with the cationic chains of the brushes. Dynamic Light Scattering is used to study the influence of the metal ions on the brushes. It is observed that metal ions induce much more pronounced shrinking as compared to the monovalent ions. The shrinking can be compared with the shrinking caused multivalent ions such as MgSO4. The metal ions once introduced are localized within the brushes due to strong correlation of counterions with the polyelectrolyte chains. The excess ions are cleaned by ultrafiltration. The trapped counterions render high contrast to the brushes and hence visualize brushes in Cryo-TEM. The metal ions can be reduced chemically by NaBH4 to generate nanoparticles. The particles are studied by high-resolution Transmission microscopy and cryogenic TEM. The metal nanoparticles formed are well spaced and crystalline in nature. The particles formed are found to be stable against aggregation. The silver nanoparticles are formed by using the anionic brushes (polyacrylic acid) and AgNO3 as the precursor salt. The catalytic activity of Platinum is tested by the heterogeneous hydrogenation of the carbonyl functionalities with dihydrogen under optimum conditions. The platinum nanoparticles give a 90% conversion of butyraldehyde to 1-butanol. The catalyst is found to be recyclable for a number of runs without losing the efficiency. The time dependent studies are carried out to gain insight in the mechanism and the kinetics of the reaction. The particles are found to be stable after the catalytic cycles. The gold nanoparticles supported on the polystyrene are found to be catalytically active for the same reaction. The gold particles are found to be recyclable for a number of runs without losing the efficiency. The bulk gold is completely inactive, hence the origin of catalytic activity is attributed to the quantum size effects. As the polystyrene particles are inert, the role of support particles in the origin of catalytic activity can be ruled out. The nanoparticles are found to become inactive for catalysis after 6 months of standing with a manifold increase in the particle size as revealed by microscopy.
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Counterion Distribution around a Macroion in Polyelectrolytes Probed by Anomalous Small-Angle X-ray Scattering
(2005)
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Mushtaq Patel
- A systematic and comprehensive study of polyelectrolytes in solution has been carried out using Anomalous Small Angle X-ray Scattering (ASAXS) in order to probe the counterion correlation around the macroion. In the course of this study, different polyelectrolyte systems such as, rod-like polyelectrolytes, spherical polyelectrolyte brushes and star shaped polyelectrolytes have been studied using ASAXS. ASAXS is the method of choice for the study of aqueous polyelectrolytes because it obviates the need of using labelled compounds or counterion exchange, as in the case of SANS or conventional SAXS. Scattering experiments performed near the absorption edge of the element under investigation, allow us to obtain the three partial scattering terms predicted by theory. All the three partial scattering terms are obtained for the first time experimentally and compared with the theoretical predictions of rod-like polyelectrolytes and spherical polyelectrolyte brushes. Rod-like polyelectrolytes bearing a rigid poly(p-phenylene) and comprising of two counterions per monomer unit with a persistence length of 21 nm were studied using ASAXS and osmometry. The results of the ASAXS experiments carried out on rod-like polyelectrolytes are compared with the predictions of the Poisson-Boltzmann Cell Model. The results suggest that the counterions are strongly correlated with the macroion. Osmotic coefficient measurements on rod-like polyelectrolytes show that approximately 20 percent of the counterions are osmotically active. The comparison of the experimentally obtained osmotic coefficient with the theory proceeds without adjusting the charge parameter. The results are in semi-quantitative agreement with the predictions of the Poisson-Boltzmann Cell Model. Spherical polyelectrolyte brushes consisting of a poly(styrene) core of 68 nm with densely grafted linear poly(acrylic acid) chains with Rubidium counterions were studied using ASAXS. In this thesis the first complete analysis of a colloidal polyelectrolyte brush by ASAXS has been presented. It has been demonstrated that there is a strong correlation of the counterions to the grafted polyelectrolyte chains of the spherical polyelectrolyte brushes. The correlation between ions and polymer chains in spherical polyelectrolyte brushes is much stronger than linear polyelectrolytes. The experiments therefore validate the theoretical predictions that most of the counterions are fully trapped within the brush and this leads to a strong stretching of the polyelectrolyte chains. Star-shaped polyelectrolytes bearing 21 arms of poly(acrylic acid) with Rubidium counterions were studied using SAXS and ASAXS. In SAXS experiments, the dependence of the maxima in the scattering experiments was observed to be concentration dependent. The ordering in star-shaped polyelectrolytes was maximum in the vicinity of the critical concentration. The maxima observed in the scattering experiments obey the scaling theory for polyelectrolytes. This is an indication of some order between the star-shaped polyelectrolytes. Evidence for the presence of any crystalline structure was not found. Scattering experiments with varying number of arms may be helpful to determine their role in the formation of crystalline structure. The three scattering terms predicted by ASAXS theory were also obtained for the star-shaped polyelectrolytes. It has been demonstrated that ASAXS is indeed a very robust method to study the polyelectrolytes in solution and to obtain the information regarding the distribution of counterions in solution. ASAXS studies of polyelectrolytes have demonstrated that the counterions are strongly correlated to the macroion.
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Visualization, Kinetics, and Thermodynamics of DNA-Protein Interactions
(2005)
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Frank Schubert
- In this work the two spectroscopic techniques surface plasmon resonance (SPR) and fluorescence correlation spectroscopy (FCS) as well as the imaging technique cryo-transmission electron microscopy (cryo-TEM) were used to gain kinetic, thermodynamic and structural information about DNA–protein interactions. Furthermore, the micrographs obtained by cryo-TEM were compared to AFM images taken in a previous work. The main goal of this work was to investigate the influence of surfaces on DNA–protein interactions, therefore the methods mentioned above were chosen. Both SPR and AFM deal with molecules attached to a surface, whereas FCS and cryo-TEM monitor the molecules in free solution. As a suitable model system the well characterized interaction between the human replication protein A (RPA) and DNA was chosen. The application of SPR and FCS to analysing the binding of RPA to ssDNA yields information about the kinetics and thermodynamics. No modification of the protein is required and biotinylated and fluorescently labelled DNA strands are available from commercial sources. Salt concentration, pH and temperature can be varied over a wide range. To best of our knowledge, FCS has not been used previously to obtain equilibrium constants at different temperatures. In this work it was demonstrated how temperature dependent SPR and FCS measurements can be performed and evaluated to determine thermodynamic data of DNA–protein interactions. Astonishingly, the equilibrium constant KD for the binding of RPA to ssDNA obtained by FCS is larger than the value obtained by SPR by a factor of 20–25, depending on the temperature. Therefore the values found for the Gibbs free energy were different, whereas the values for the reaction enthalpy were nearly the same for the two methods used. There are clear evidences that the difference in KD and therefore in Gibbs free energy measured by the two methods is due to different reaction entropies. In SPR the reaction is restricted to two dimensions due to immobilization of the DNA molecules to the sensor surface, thus the rate constants obtained might not be the true association and dissociation rates. As a main result, the data obtained by SPR differ from the data gained from the free solution experiments. The reason for this is a loss of one degree of freedom, which in turn results in different entropic terms for the surface and the free solution techniques. In contrast, FCS is able to follow complex formation without spatial restrictions. In consequence, the reaction in three dimensions is entropically less favourable than the reaction at the solid-liquid interface. This might be due to differences in the cratic entropy between the two geometries, however, the role of hydration can not be assessed by our experiments. The picture of the DNA–RPA interaction was completed by further FCS measurements using various dsDNA fragments containing damage sites. The binding of RPA to undamaged dsDNA fragments showed a low affinity to dsDNA (approx. 15%), as expected from previous AFM experiments. Since RPA is known to have a high affinity to singlestranded DNA, this finding may be explained by the binding of RPA to unpaired nucleotides at the end of the dsDNA. Comparing the two imaging techniques AFM and cryo-TEM one does not find a strong influence of the surface on the DNA–RPA interaction. The kinks formed by UV-damaged DNA observed in AFM experiments could not be verified by the cryo-TEM experiments. There might be two reasons for this: First, the kinks in the AFM experiments are induced by the mica surface and therefore do not occur in cryo-TEM experiments. Second, the resolution of the TEM is not as good as in AFM, therefore, the kinks can not be seen in the TEM. The question if the DNA is wrapping around the RPA as stated in earlier works can not be answered using cryo-TEM. The resolution of this method is not as good as in AFM. In order to get micrographs with a better resolution one has to perform simple TEM experiments including staining of the molecules. The drawback of this procedure is that the molecules are influenced by the staining chemicals and therefore not in their natural state. In a very last part of this work the mini-chromosome maintenance com-plex was investigated using FCS and cryo-TEM. It was shown that the protein exhibits a medium affinity to ssDNA and dsDNA. The structure of the DNA substrate does not play an important role, the interaction was the same for simple and bubble dsDNA and dsDNA containing a ssDNA tail.
