Untersuchung einzelner SNARE-vermittelter Membranfusionsereignisse auf planaren porenüberspannenden Membranen

Schwenen, Lando Lantbert Gregor

dc.contributor.advisor	Steinem, Claudia Prof. Dr.
dc.contributor.author	Schwenen, Lando Lantbert Gregor
dc.date.accessioned	2015-10-20T09:13:28Z
dc.date.available	2015-10-20T09:13:28Z
dc.date.issued	2015-10-20
dc.identifier.uri	http://hdl.handle.net/11858/00-1735-0000-0023-9650-F
dc.identifier.uri	http://dx.doi.org/10.53846/goediss-5308
dc.language.iso	deu	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	Untersuchung einzelner SNARE-vermittelter Membranfusionsereignisse auf planaren porenüberspannenden Membranen	de
dc.type	doctoralThesis	de
dc.title.translated	Investigation of Single SNARE-mediated Membrane Fusion Events on Planar Pore-spanning Membranes	de
dc.contributor.referee	Steinem, Claudia Prof. Dr.
dc.date.examination	2015-06-04
dc.description.abstracteng	Even though there are a number of diﬀerent in vitro fusion assays to analyze neuronal SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) mediated fusion, still the essential features of the in vivo situation are only partially captured. In this thesis an in vitro fusion assay that provides a planar solvent-free pore-spanning membrane containing a ternary acceptor SNARE complex, which mimics the presynaptic membrane, to monitor fusion of single synaptobrevin containing vesicles is presented. Protein-containing pore-spanning membranes were obtained by spreading of giant unilamellar vesicles on gold covered6-mercapto-1-hexanol functionalized porous siliconnitride substrates with pore diameters of 1.2µm. The mobility of Lipids and the SNAREs in the pore-spanning membranes was shown by FCS-experiments, which is prerequisite for the formation of active fusion complexes. The fusion process was highly speciﬁc with an eﬃciency of 50 % and was analyzed by two color confocal laser scanning ﬂuorescence microscopy in a timeresolved manner allowing to distinguish between vesicle docking, hemifusion and full fusion. A kinetics analysis revealed that two reaction steps need to take place to progress to fusion with a lifetime of the docked vesicles of about 50s.	de
dc.contributor.coReferee	Salditt, Tim Prof. Dr.
dc.subject.ger	SNARE	de
dc.subject.ger	Membranfusion	de
dc.subject.ger	Fusionsassay	de
dc.subject.ger	porenüberspannende Membran	de
dc.subject.ger	Diffusion in Lipidmembranen	de
dc.subject.ger	konfokale Laserrastermikroskopie	de
dc.subject.ger	Hemifusion	de
dc.subject.ger	Oberflächenfunktionalisierung	de
dc.subject.ger	poröse Substrate	de
dc.subject.ger	Lipidmembranen	de
dc.subject.ger	Vesikelfusion	de
dc.subject.ger	Mobilität von SNARE-Proteinen	de
dc.subject.eng	SNARE	de
dc.subject.eng	soluble N-ethylmaleimide-sensitive factor attachment protein receptor	de
dc.subject.eng	fusion	de
dc.subject.eng	fusion assay	de
dc.subject.eng	pore-spanning membranes	de
dc.subject.eng	mobility of Lipids	de
dc.subject.eng	mobility of SNAREs	de
dc.subject.eng	porous substrates	de
dc.subject.eng	vesicle docking	de
dc.subject.eng	hemifusion	de
dc.subject.eng	confocal laser scanning ﬂuorescence microscopy	de
dc.subject.eng	lipid membranes	de
dc.subject.eng	surface functionalization	de
dc.subject.eng	membrane fusion	de
dc.subject.eng	membrane diffusion	de
dc.identifier.urn	urn:nbn:de:gbv:7-11858/00-1735-0000-0023-9650-F-2
dc.affiliation.institute	Fakultät für Chemie	de
dc.subject.gokfull	Chemie (PPN62138352X)	de
dc.identifier.ppn	837584302

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