Kinetic analysis of neuronal SNARE protein interactions

Pribićević, Sonja

dc.contributor.advisor	Jahn, Reinhard Prof. Dr.
dc.contributor.author	Pribićević, Sonja
dc.date.accessioned	2022-08-08T12:39:45Z
dc.date.available	2022-11-01T00:50:08Z
dc.date.issued	2022-08-08
dc.identifier.uri	http://resolver.sub.uni-goettingen.de/purl?ediss-11858/14202
dc.identifier.uri	http://dx.doi.org/10.53846/goediss-9392
dc.language.iso	eng	de
dc.rights.uri	http://creativecommons.org/licenses/by/4.0/
dc.subject.ddc	572	de
dc.title	Kinetic analysis of neuronal SNARE protein interactions	de
dc.type	doctoralThesis	de
dc.contributor.referee	Jahn, Reinhard Prof. Dr.
dc.date.examination	2021-11-03	de
dc.description.abstracteng	When an action potential arrives at the axon terminal, it causes a local and transient increase in the Ca2+ concentration that starts a cascade of molecular interactions eventually leading to the fusion of synaptic vesicles with the plasma membrane. The key proteins in this process – syntaxin-1A, SNAP25 and synaptobrevin 2 – assemble into a SNARE complex that directly drives the membrane fusion. The interaction of the SNARE proteins and mechanism by which this interaction mediates membrane fusion is intensively investigated. Despite numerous studies, the answers to the core questions regarding the molecular mechanism, the guidance and the speed of SNARE complex formation remain unclear. In this work, I address one of these core questions, namely, the molecular mechanism of SNARE assembly as it represents a fundamental problem that underlies the understanding of any other process regarding SNARE-mediated fusion. Using purified, soluble SNARE proteins and kinetic measurements under different conditions, I identified some of the most likely intermediates of the SNARE assembly reaction and estimated the rate constants of their formation. Accordingly, I confirmed that the syntaxin-1A-SNAP25 interaction represents the first of the SNARE complex assembly reaction. Size exclusion chromatography experiments provided further details on the possible intermediates by showing that stable complexes can form between syntaxin-1A, synaptobrevin 2 and the individual SNARE domains of SNAP25. I conclude that SNARE complex formation occurs in at least two steps, and is initiated by the interaction between syntaxin-1A and SNAP25. The results and the experimental approach developed in this work represent a backbone for elucidating the complicated interactions that control synaptic vesicle fusion.	de
dc.contributor.coReferee	Moser, Tobias Prof. Dr.
dc.contributor.thirdReferee	Rodnina, Marina Prof. Dr.
dc.subject.eng	SNARE	de
dc.subject.eng	kinetics	de
dc.subject.eng	syntaxin-1A	de
dc.subject.eng	SNAP25	de
dc.subject.eng	synaptobrevin 2	de
dc.subject.eng	FRET	de
dc.subject.eng	stopped-flow	de
dc.identifier.urn	urn:nbn:de:gbv:7-ediss-14202-9
dc.affiliation.institute	Göttinger Graduiertenschule für Neurowissenschaften, Biophysik und molekulare Biowissenschaften (GGNB)	de
dc.subject.gokfull	Biologie (PPN619462639)	de
dc.description.embargoed	2022-11-01	de
dc.identifier.ppn	181392497X
dc.notes.confirmationsent	Confirmation sent 2022-08-08T12:45:01	de

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GGNB - Göttinger Graduiertenzentrum für Neurowissenschaften, Biophysik und molekulare Biowissenschaften [1112]
GGNB - Göttingen Graduate Center for Neurosciences, Biophysics and Molecular Biosciences

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