On the molecular basis of α-synuclein aggregation  on phospholipid membranes in the presence and  absence of anle138b

Antonschmidt, Leif

dc.contributor.advisor	Griesinger, Christian Prof. Dr.
dc.contributor.author	Antonschmidt, Leif
dc.date.accessioned	2020-07-22T13:41:49Z
dc.date.available	2021-11-27T00:50:03Z
dc.date.issued	2020-07-22
dc.identifier.uri	http://hdl.handle.net/21.11130/00-1735-0000-0005-1428-8
dc.identifier.uri	http://dx.doi.org/10.53846/goediss-8097
dc.language.iso	eng	de
dc.rights.uri	http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.ddc	571.4	de
dc.title	On the molecular basis of α-synuclein aggregation on phospholipid membranes in the presence and absence of anle138b	de
dc.type	doctoralThesis	de
dc.title.translated	Zur molekularen Basis der α-Synuclein Aggregation an Phospholipid Membranen in der Gegenwart und Abwesenheit von anle138b
dc.contributor.referee	Griesinger, Christian Prof. Dr.
dc.date.examination	2019-11-28
dc.description.abstracteng	A physiologic hallmark of synucleinopathies such as Parkinson’s Disease (PD) or Multiple System Atrophy (MSA) are deposits in patients’ brains, which consist of the α-synuclein. Growing evidence suggests, that oligomeric protein intermediates, formed during the aggregation process, constitute the major toxic species. The 3,5-diphenyl-pyrazole derivative anle138b has been designed and synthesized to interfere with these intermediates and showed significant efficacy in mouse models of Parkinson’s Disease. The success of anle138b in vivo calls for an elucidation of the underlying mechanism in vitro. In this thesis a combination of NMR-, fluorescence- and CD-spectroscopy was used to thoroughly characterize the kinetics of α-synuclein (αS) in the presence of phospholipid membranes under Protein Misfolding Cyclic Amplification (PMCA) conditions. The exact knowledge of the kinetics in return allowed the enrichment of oligomeric αS intermediates. Morphological information on these intermediates was obtained by AFM-microscopy and Transmission Electron Microscopy. Solid-sate NMR revealed that formation of two loop regions in these pre-organized intermediate initiates αS fibril formation. This species then rapidly rearranges with parts of the NAC region adopting the final fibril conformation. The structural changes observed during aggregation are each accompanied by a distinct response to ThT, allowing identification of key time points for the isolation of intermediates. Using DNP-enhanced solid-state-NMR in combination with photoaffinity studies an interaction of membrane-embedded anle138b with αS oligomers and fibrils was characterized. Site specific information was refined by MD-simulations for αS fibrils. In conclusion, the findings in this work add to the understanding of both the mechanism of a αS aggregation and the interaction of aggregates with small molecules.	de
dc.contributor.coReferee	Steinem, Claudia Prof. Dr.
dc.subject.eng	amyloid	de
dc.subject.eng	alpha-synuclein	de
dc.subject.eng	protein-small molecule interaction	de
dc.subject.eng	phospholipid bilayers	de
dc.subject.eng	PMCA	de
dc.subject.eng	aggregation intermediates	de
dc.subject.eng	membrane insertion	de
dc.subject.eng	BSA	de
dc.subject.eng	aggregation mechanism	de
dc.subject.eng	oligomers	de
dc.subject.eng	aggregation kinetics	de
dc.identifier.urn	urn:nbn:de:gbv:7-21.11130/00-1735-0000-0005-1428-8-7
dc.affiliation.institute	Göttinger Graduiertenschule für Neurowissenschaften, Biophysik und molekulare Biowissenschaften (GGNB)	de
dc.subject.gokfull	Biologie (PPN619462639)	de
dc.description.embargoed	2020-11-26
dc.description.embargoed	2021-11-27
dc.identifier.ppn	1725349124

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GGNB - Göttinger Graduiertenzentrum für Neurowissenschaften, Biophysik und molekulare Biowissenschaften [1115]
GGNB - Göttingen Graduate Center for Neurosciences, Biophysics and Molecular Biosciences

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