Degree Type

Dissertation

Date of Award

2008

Degree Name

Doctor of Philosophy

Department

Biochemistry, Biophysics and Molecular Biology

First Advisor

Yeon-Kyun Shin

Second Advisor

Alan Disprito

Third Advisor

Edward Yu

Abstract

At the synapse, neurotransmitters are released via Ca2+-triggered exocytotic fusion of synaptic vesicles with the presynaptic plasma membrane. The whole process is controlled by various proteins. SNAREs have been recognized as the key components that drive membrane fusion. In addition, many other proteins, such as Munc18/nSec1, Mun13, synaptotagmin, complexin, etc. are characterized to regulate synaptic transmission temporally and spatially.;The in vitro bulk fluorescence assay was applied to examine the kinetics of membrane fusion of liposomes mediated by recombinant neuronal SNAREs and led to the demonstration of hemifusion as an intermediate in the pathway. In order to monitor the fusion process in more sophisticated level, we developed the Single-liposome FRET assay and combined site-directed spin labeling (SDSL) and electron paramagnetic resonance (EPR) techniques to study the function of two SNARE regulators, synaptotagmin and complexin.;The EPR and fluorescence assay were also applied for the study of the SNAREs mediating the trafficking in yeast. It was found that supermolecular SNARE assembly precedes hemifusion, which was subsequently followed by distal leaflet mixing and formation of the cis-SNARE complex.

DOI

https://doi.org/10.31274/rtd-180813-17016

Publisher

Digital Repository @ Iowa State University, http://lib.dr.iastate.edu/

Copyright Owner

Xiaobing Lu

Language

en

Proquest ID

AAI3316168

OCLC Number

271241014

ISBN

9780549687856

File Format

application/pdf

File Size

145 pages

Included in

Biophysics Commons

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