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

Groth, Mike Christopher

dc.contributor.advisor	Diederichsen, Ulf Prof. Dr.
dc.contributor.author	Groth, Mike Christopher
dc.date.accessioned	2021-03-09T12:22:00Z
dc.date.available	2021-03-16T23:50:03Z
dc.date.issued	2021-03-09
dc.identifier.uri	http://hdl.handle.net/21.11130/00-1735-0000-0005-159B-5
dc.identifier.uri	http://dx.doi.org/10.53846/goediss-8480
dc.identifier.uri	http://dx.doi.org/10.53846/goediss-8480
dc.language.iso	eng	de
dc.publisher	Niedersächsische Staats- und Universitätsbibliothek Göttingen	de
dc.rights.uri	http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.ddc	540	de
dc.title	Investigation of the Linker Region of Coiled Coil SNARE-Analoga and Membrane Composition on Vesicle Fusion	de
dc.type	doctoralThesis	de
dc.contributor.referee	Diederichsen, Ulf Prof. Dr.
dc.date.examination	2021-01-11
dc.description.abstracteng	SNARE (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 events.	de
dc.contributor.coReferee	Steinem, Claudia Prof. Dr.
dc.subject.eng	SNARE	de
dc.subject.eng	coiled coil	de
dc.subject.eng	Fluorescence Spectroscopy	de
dc.subject.eng	Vesicle fusion assays	de
dc.subject.eng	Dynamic light scattering (DLS)	de
dc.subject.eng	lipid membrane	de
dc.subject.eng	photo cage	de
dc.subject.eng	fluorescence resonance energy transfer (FRET)	de
dc.identifier.urn	urn:nbn:de:gbv:7-21.11130/00-1735-0000-0005-159B-5-2
dc.affiliation.institute	Fakultät für Chemie	de
dc.subject.gokfull	Chemie (PPN62138352X)	de
dc.description.embargoed	2021-03-16
dc.identifier.ppn	1750898888

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Fakultät für Chemie (inkl. GAUSS) [701]

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