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Investigation of the Linker Region of Coiled Coil SNARE-Analoga and Membrane Composition on Vesicle Fusion

dc.contributor.advisorDiederichsen, Ulf Prof. Dr.
dc.contributor.authorGroth, Mike Christopher
dc.publisherNiedersächsische Staats- und Universitätsbibliothek Göttingende
dc.titleInvestigation of the Linker Region of Coiled Coil SNARE-Analoga and Membrane Composition on Vesicle Fusionde
dc.contributor.refereeDiederichsen, Ulf Prof. Dr.
dc.description.abstractengSNARE (soluble N-ethylmaleimide-sensitive-factor attachment receptor) proteins play a major role in membrane fusion processes in many organisms. As these proteins form a sophisticated tetrameric coiled coil complex, many artificial models have been developed to mimic the natural system and reduce its complexity. The model system used in this work uses a combination of a coiled-coil (E3/K3) pair as the recognition unit and the native sequence of syntaxin 1A and synaptobrevin 2 for the linker and transmembrane region. These two peptides form a stable dimer upon interaction with each other and are capable of performing membrane fusion, as it has been shown previously. In this thesis, improvements of the peptide synthesis, purification and employed fusion assays have been established. DLS measurements have been introduced as a reliable technique for observing vesicle quality. Additionally, this work focusses on the effects on the fusion efficiency of specific amino acids of the linker domains. Strategic substitution of charged and aromatic amino acids showed a decrease in the fusion activity of the system. Furthermore, mechanistical studies were performed targeting the role of the linker domain by the introduction of a sterically demanding photo cage. Fusion activity was reduced by the photo cage, but could be successfully restored after the photo cleaving process. In addition, introduction of charged lipids (PIP2, DOPS) into the vesicle membranes lead to a complete stop of fusion
dc.contributor.coRefereeSteinem, Claudia Prof. Dr.
dc.subject.engcoiled coilde
dc.subject.engFluorescence Spectroscopyde
dc.subject.engVesicle fusion assaysde
dc.subject.engDynamic light scattering (DLS)de
dc.subject.englipid membranede
dc.subject.engphoto cagede
dc.subject.engfluorescence resonance energy transfer (FRET)de
dc.affiliation.instituteFakultät für Chemiede
dc.subject.gokfullChemie  (PPN62138352X)de

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