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@PHDTHESIS{vanMaarschalkerweerd:206202,
author = {van Maarschalkerweerd, Andreas},
title = {{S}tructural and biophysical characterization of
$\alpha$-synuclein and itsinteraction with lipid membranes},
school = {University of Copenhagen},
type = {Dr.},
reportid = {PUBDB-2015-00716},
pages = {-},
year = {2014},
note = {Post referee full text in progress.; University of
Copenhagen, Diss., 2014},
abstract = {Parkinson's disease (PD) is a pervasive neurodegenerative
disorder affecting millions of lives worldwide. There is
currently no cure for PD and the pharmacological treatment
is hence only symptomatic. Despite immense scientific
efforts, there is still a lack of basic understanding of the
pathogenesis of PD. Such understanding might be the key to
optimized drug discovery strategies. Due to the outspoken
similarities between PD and a number of other
neurodegenerative and amyloid diseases, an enhanced
understanding of PD is potentially also of general
interest.PD is hallmarked by the presence of lipid-rich
protein inclusions in the brain called Lewy Bodies (LB). LB
are characterized by a high content of the
$\alpha$-synuclein ($\alpha$SN) protein. A number of
familiar point mutations within $\alpha$SN have been shown
to cause early-onset genetic PD. The native structure of
αSN is generally believed to be intrinsically disordered.
However, within LB $\alpha$SN is found as elongated
aggregates (amyloid fibrils) characterized by cross-$\beta$
structure. The fibrillated $\alpha$SN found in LB is
believed to be an inert reservoir. The cytotoxic effect of
$\alpha$SN is allegedly associated with the occurrence of
transient oligomer structures that form prior to the
formation of amyloid fibrils. Several studies have sought to
elucidate on the molecular mechanism of $\alpha$SN
interaction with lipid membranes in relation to the apparent
cytotoxicity of oligomeric $\alpha$SN.This thesis presents
experimental work addressing the structure of $\alpha$SN,
aspects of $\alpha$SN-membrane interaction and process
behind $\alpha$SN mutant fibrillation. The primary
experimental methods are small-angle X-ray scattering (SAXS)
and fluorescence spectroscopy with emphasis on the
environmental sensitive dye Laurdan.During purification of
$\alpha$SN we observed an unexpected chromatographic
feature. This feature was revealed to be a consequence of
the formation of partly compacted covalent αSN dimers as
shown by e.g. SAXS and mass spectroscopy. Two projects were
performed on the interactions of $\alpha$SN and lipid
membranes. Here we showed that $\alpha$SN is able to
dehydrate anionic membranes and to induce lysis of liposomes
associated with the formation of $\alpha$-helical rich
$\alpha$SN-lipid co-aggregates. Additionally we showed that
cholesterol could modulate the interaction between
$\alpha$SN oligomers and otherwise inert neutral lipid
membranes. Finally a SAXS based study was conducted on the
fibrillation of $\alpha$SN E46K mutant proving that the
fibrillation of E46K is significantly different from that of
wild-type $\alpha$SN under the applied conditions.},
keywords = {Dissertation (GND)},
cin = {EMBL-User},
cid = {I:(DE-H253)EMBL-User-20120814},
pnm = {DORIS Beamline D1.2 (POF2-54G13) / PETRA Beamline P12
(POF2-54G14)},
pid = {G:(DE-H253)POF2-D1.2-20130405 /
G:(DE-H253)POF2-P12-20130405},
experiment = {EXP:(DE-H253)D-D1.2-20150101 / EXP:(DE-H253)P-P12-20150101},
typ = {PUB:(DE-HGF)11},
url = {https://bib-pubdb1.desy.de/record/206202},
}