The ubiquitination of proteins serves as molecular signal to control an enormous number of physiological processes and its dysregulation is connected to human diseases like cancer. The versatility of this signal stems from the diverse ways by which ubiquitin can be attached to its targets. Thus, specificity and tight regulation of the ubiquitination are pivotal requirements of ubiquitin signaling. Ubiquitin-conjugating enzymes (E2s) act at the heart of the ubiquitination cascade, transferring ubiquitin from a ubiquitin-activating enzyme (E1) to a ubiquitin ligase (E3) or substrate. When cooperating with a RING-type E3, ubiquitin-conjugating enzymes can determine linkage specificity in ubiquitin chain formation. Our understanding of the regulation of E2 activities is still limited at a structural level. The work described here identifies two regulation mechanisms in UBE2S, a cognate E2 of the human RING-type E3 anaphase-promoting complex/cyclosome (APC/C). UBE2S elongates ubiquitin chains on APC/C substrates in a Lys11 linkage-specific manner, thereby targeting these substrates for degradation and driving mitotic progression. In addition, UBE2S was found to have a role in DNA repair by enhancing non-homologous end-joining (NHEJ) and causing transcriptional arrest at DNA damage sites in homologous recombination (HR). Furthermore, UBE2S overexpression is a characteristic feature of many cancer types and is connected to poor prognosis and diminished response to therapy. The first regulatory mechanism uncovered in this thesis involves the intramolecular auto-ubiquitination of a particular lysine residue (Lys+5) close to the active site cysteine, presumably through conformational flexibility of the active site region. The Lys+5-linked ubiquitin molecule adopts a donor-like, ‘closed’ orientation towards UBE2S, thereby conferring auto-inhibition. Notably, Lys+5 is a major physiological ubiquitination site in ~25% of the human E2 enzymes, thus providing regulatory opportunities beyond UBE2S. Besides the active, monomeric state and the auto-inhibited state caused by auto-ubiquitination, I discovered that UBE2S can adopt a dimeric state. The latter also provides an auto-inhibited state, in which ubiquitin transfer is blocked via the obstruction of donor binding. UBE2S dimerization is promoted by its unique C-terminal extension, suppresses auto-ubiquitination and thereby the proteasomal degradation of UBE2S. Taken together, the data provided in this thesis illustrate the intricate ways by which UBE2S activity is fine-tuned and the notion that structurally diverse mechanisms have evolved to restrict the first step in the catalytic cycle of E2 enzymes. / Die Ubiquitinierung von Proteinen fungiert als molekulares Signal zur Kontrolle einer Vielzahl physiologischer Prozesse, wobei eine gestörte Regulation der Ubiquitinierung eng mit zahlreichen Erkrankungen, wie beispielsweise Krebs, verbunden ist. Aufgrund der verschiedenen Verknüpfungsmöglichkeiten von Ubiquitin, die das zelluläre Schicksal des Zielproteins bestimmen, sind Spezifität und stringente Regulation unabkömmliche Voraussetzungen im Ubiquitinierungsprozess. Ubiquitin-konjugierende Enzyme (E2s) fungieren in der Mitte der Ubiquitinierungskaskade. Sie übernehmen ein Ubiquitinmolekül vom Ubiquitin-aktivierenden Enzym (E1) und übertragen es auf eine Ubiquitin-Ligase (E3) oder direkt auf das Zielprotein. Arbeiten Ubiquitin-konjugierende Enzyme mit E3s des RING-Typus zusammen, so bestimmen E2s die Art der Verknüpfung. Die Regulation der Aktivität Ubiquitin-konjugierender Enzyme auf struktureller Ebene ist jedoch bisher nur bedingt verstanden. Die hier dargelegte Arbeit umfasst die Identifizierung zweier Regulationsmechanismen des Ubiquitin-konjugierenden Enzyms UBE2S. UBE2S arbeitet mit einem humanen E3 des RING-Typus‚ dem ‚Anaphase Promoting Complex/Cyclosome‘ (APC/C) zusammen und bildet Lys11-spezifische Ubiquitinketten auf Substraten des APC/Cs. Hierdurch werden die Substrate für den Abbau durch das Proteasom markiert, was das Fortschreiten der Mitose bedingt. Zusätzlich wird UBE2S eine Rolle in der DNS-Reparatur zugeschrieben. Hierbei verstärkt UBE2S die nicht-homologe Rekombination (NHEJ) und verhindert außerdem die Transkription an DNS-Bruchstellen, die durch Homologe Rekombination (HR) repariert werden. Die Überexpression von UBE2S ist ein Charakteristikum verschiedenster Krebsarten, vermindert den Erfolg herkömmlicher Krebstherapien, und führt somit zu schlechten Prognosen für betroffenen Patienten. Der erste hier beschriebene Regulationsmechanismus beinhaltet die intramolekulare Ubiquitinierung eines Lysins (Lys+5) nahe des katalytischen Cysteins, mutmaßlich durch strukturelle Flexibilität der Region des aktiven Zentrums. Das Lys+5-verknüpfte Ubiquitin nimmt eine Donorubiquitin-ähnliche Position auf UBE2S ein, wodurch UBE2S gehemmt wird. Da ein Lysin an der Position +5 in ~25% der humanen E2-Enzyme vorhanden und eine physiologische Ubiquitinierungsstelle ist, birgt dieser Mechanismus Regulationsmöglichkeiten über UBE2S hinaus. Zusätzlich zum aktiven monomeren Zustand und dem durch Autoubiquitinierung ausgelösten inhibierten Zustand, kann UBE2S auch als Dimer vorliegen. In diesem Zustand ist es ebenfalls inaktiv, da die Donorubiquitin-Bindestelle auf UBE2S durch ein zweites Molekül des E2s blockiert wird. Begünstigt wird die Dimerisierung durch die C-terminale Verlängerung von UBE2S und verhindert so deren Autoubiquitinierung, und folglich den proteasomalen Abbau von UBE2S. Es handelt sich hierbei somit um einen zweiten Regulationsmechanismus von UBE2S. Zusammenfassend veranschaulichen die in dieser Arbeit dargelegten Daten die komplexen Möglichkeiten, durch die die Aktivität von UBE2S reguliert werden kann, sowie die Erkenntnis, dass strukturell unterschiedliche Mechanismen existieren, um den ersten Schritt der von Ubiquitin-konjugierenden Enzymen katalysierten Reaktion zu hemmen.

This master´s thesis deals with a proposed design of the derrick of the filling car of a coke-oven battery, which is used for working with unit loads, repairing and maintenance of the filling car and cleaning of the battery risers. In the first part a solution concept was chosen. For this concept a main steel structure was designed, which was controlled in stress analysis by the FEM program I-DEAS NX 11. Further, the main parts of the lifting and travelling mechanism were calculated using specified parameters. In the conclusion of this thesis the results summary of specified targets is stated. Assembly drawing is in the annex of this thesis.

The diploma thesis deals with some parts of bridge magnet crane with 15 tons of nominal lifting capacity. This crane is designed for operation in the steel mill for handling billets using magnets suspended on the traverse. This work is focused on design of hoisting and traverse mechanism of crane trolley, selection of individual components on the basis of relevant standards and calculations. Next part of this thesis is investigation magnet traverse in terms of its strength according to standard ČSN EN 13001.

Diplomová práce popisuje vývoj objemu a struktury poskytovaných úvěrů v ČR v oblasti veřejného a podnikového zadlužení se zaměřením na zadlužení v segmentu domácností. Zde je provedena analýza faktorů, které ovlivňují změny ve struktuře a dynamice úvěrů. Významný vliv má vývoj úrokových sazeb, změny úvěrových podmínek, růst disponibilních příjmů, trh nemovitostí či nové trendy v přístupu k zadlužování.

Zugl.: Giessen, Univ., Diss., 2008

University, Diss., 2004--Heidelberg.

This thesis examines the sensitivity of bank interest rates to changes in in-terest rates CNB's monetary policy in the context of interest rate transmission mechanism. For this reason, emphasis was focused on the functioning of a basic form of transmission mechanism and theoretical approaches to economic trends . Further determinants of bank interest rates were indetified and further described the issue of heterogeneity in the transmission of interest rates. In the practical part the author of the work focused on examining the sensitivity with which bank in adjusting their interest rates on newly granted mortgage loans to changes in short-term interest rates in the interbank market. The observed heterogeneity was further examined in relation to the factors of concentration, market power, liquidity, capital adequacy and credit risks that have been identified on the basis of empirical studies Sorenson and Werner (2006 ). With the help of correlation analysis it was concluded that banks react heterogeneous manner even within homogeneous groups where banks have a large degree of simi-larity.

Polychlorinated biphenyls (PCBs) are chlorinated organic compounds, which belong to persistent organic pollutants and exhibit various modes of toxic action, including mutagenicity, carcinogenicity, and endocrine disruption. PCBs were manufactured during the 20th century in many countries and extensively used due to their advantageous physicochemical properties. PCBs mostly served as insulating liquids in electrical equipment; however, they were also utilized in many open applications. Despite the worldwide ban on PCB manufacture imposed at the end of the 20th century, the contamination of the environment persists to this day as a result of their recalcitrance. Moreover, PCBs are still being inadvertently produced during many industrial activities. Because of their stability, the breakdown of PCBs in nature is extremely slow. This dissertation thesis focuses on the study of PCB biodegradation by ligninolytic fungi. This group of microorganisms belongs to the most promising, especially in regard to the degradation of organic pollutants. The biodegradation mechanism of PCBs was studied in vitro, including the identification of degradation intermediates. Laccase, an enzyme expressed by the oyster mushroom (strain Pleurotus ostreatus 3004), was able to degrade hydroxylated PCBs. In addition, chlorinated...

This master’s thesis deals with the piston for modified engine 1.4 TSI from Volkswagen concern. It includes calculation of kinematics and forces reactions of the crank mechanism of the engine for speed 6071 rpm elaborated in the program Mathcad Prime. Further the work includes calculation for reducing the magnitude of lateral force and design of several variants of the piston. For the proposed design variants of the piston are performed FEM analysis of mechanical state of stress and comparisons of contact pressures in the program ANSYS Workbench.

This article contains description of basic principles of stirling engine, construction types and its parts, which supplies motor run and description of using in real life too. The main topic is analyses of driving mechanism supplying both pistons movement of stirling engine on Robert Stirling‘s drawing from 1816.