The periplasmic domain of the barrel assembly machinery protein A (BamA) from Escherichia coli assists folding of outer membrane protein A

The assembly of outer membranes of the cell wall of Gram-negative bacteria and of various organelles of eukaryotic cells requires the evolutionarily conserved β-barrel-assembly machinery (BAM) complex. This thesis describes the biochemical and biophysical properties of the periplasmic domain of the β-barrel assembly machinery protein A (PD-BamA) of the E. coli BAM complex, its effect on insertion and folding of the Outer membrane protein A (OmpA) into lipid bilayers and the identification of regions of PD-BamA that may be involved in protein-protein interactions. The secondary structure of PD-BamA in mixed lipid bilayers, analyzed by Circular dichroism (CD) spectroscopy, contained less β-sheet at an increased content of phosphatidylglycerol (PG) in the lipid membrane. This result showed membrane binding, albeit only in the presence of negatively charged lipids. Fluorescence spectroscopy demonstrated that PD-BamA only binds to lipid bilayers containing the negatively charged DOPG, confirming the results of CD spectroscopy. PD-BamA did not bind to zwitterionic but overall neutral lipid bilayers. PD-BamA bound to OmpA at a stoichiometry of 1:1. PD-BamA strongly facilitated insertion and folding of OmpA into lipid membranes. Kinetics of PD-BamA mediated folding of OmpA was well described by two parallel folding processes, a fast folding process and a slow folding process, differing by 2-3 orders of magnitude in their rate constants. The folding yields of OmpA depended on the concentration of lipid membranes and also on the lipid head groups. The presence of PD-BamA resulted in increased folding yields of OmpA in negatively charged DOPG, but PD-BamA did not affect the folding kinetics of OmpA into bilayers of zwitterionic but overall neutral lipids. The efficiency of folding and insertion of OmpA into lipid bilayers strongly depended on the ratio PD-BamA/OmpA and was optimal at equimolar concentrations of PD-BamA and OmpA. To examine complexes of unfolded OmpA with PD-BamA in more detail, site-directed spectroscopy was used to explore contact regions in both, PD-BamA and OmpA. Similarly, contact regions were also investigated for another protein complex formed by PD-BamA and the lipoprotein BamD. The obtained data suggest, that the site of interaction on PD-BamA for OmpA might be oriented towards the exterior environment away from the preceding POTRA domains, but that PD-BamA is oriented with its short α-helix α1 of POTRA domain 5 towards the C-terminal end of BamD.

Fundamental interactions of molecules (Na_2, Na_3) with intense femtosecond laser pulses

The real-time dynamics of multiphoton ionization and fragmentation of molecules Na_2 and Na_3 has been studied in molecular beam experiments employing ion and electron spectroscopy together with femtosecond pump-probe techniques. Experiments with Na_2 and Na_3 reveal unexpected features of the dynamics of the absorption of several photons as seen in the one- and three-dimensional vibrational wave packet motion in different potential surfaces and in high laser fields: In Na_2 a second major resonance-enhanced multiphoton ionization (REMPI) process is observed, involving the excitation of two electrons and subsequent electronic autoionization. The possibility of controlling a reaction by controlling the duration of propagation of a wave packet on an electronically-excited surface is demonstrated. In high laser fields, the contributions from direct photoionization and from the second REMPI process to the total ion yield change, due to different populations in the electronic states participating in the multiphoton ionization (MPI) processes. In addition, a vibrational wave packet motion in the electronic ground state is induced through stimulated emission pumping by the pump laser. The 4^1 \summe^+_g shelf state of Na_2 is given as an example for performing frequency spectroscopy of highlying electronic states in the time domain. Pure wave packet effects, such as the spreading and the revival of a vibrational wave packet, are investigated. The three-dimensional wave packet motion in the Na_3 reflects the normal modes in the X and B states, and shows in addition the pseudorotational motion in the B state in real time.

Human and Raptor Interactions in the Context of a Nomadic Society

This research project focuses on contemporary eagle-taming falconry practice of the Altaic Kazakhs animal herding society in Bayan Ulgii Province in Western Mongolia. It aims to contributing both theoretical and empirical criteria for cultural preservation of Asian falconry. This cultural as well as environmental discourse is illustrated with concentrated field research framed by ecological anthropology and ethno-ornithology from the viewpoint of “Human-Animal Interaction (HAI)” and “Human-Animal Behavior (HAB)”. Part I (Chapter 2 & 3) explores ethno-archaeological and ethno-ornithological dimensions by interpretive research of archaeological artefacts which trace the historical depth of Asian falconry culture. Part II (Chapter 4 & 5) provides an extensive ethnographic narrative of Altaic Kazakh falconry, which is the central part of this research project. The “Traditional Art and Knowledge (TAK)” in human-raptor interactions, comprising the entire cycle of capture, perch, feeding, training, hunting, and release, is presented with specific emphasis on its relation to environmental and societal context. Traditional falconry as integral part of a nomadic lifestyle has to face some critical problems nowadays which necessitate preventing the complete disappearance of this outstanding indigenous cultural heritage. Part III (Chapter 6 & 7) thus focuses on the cultural sustainability of Altaic Kazakh falconry. Changing livelihoods, sedentarisation, and decontextualisation are identified as major threats. The role of Golden Eagle Festivals is critically analysed with regard to positive and negative impact. This part also intends to contribute to the academic definition of eagle falconry as an intangible cultural heritage, and to provide scientific criteria for a preservation master plan, as well as stipulate local resilience by pointing to successive actions needed for conservation. This research project concludes that cultural sustainability of Altaic Kazakh falconry needs to be supported from the angles of three theoretical frameworks; (1) Cultural affairs for protection based on the concept of nature-guardianship in its cultural domain, (2) Sustainable development and improvement of animal herding productivity and herder’s livelihood, (3) Natural resource management, especially supporting the population of Golden Eagles, their potential prey animals, and their nesting environment.