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Surface induced Oligomerisation and Fibrillization of alpha-Synuclein investigated by 3D-SAF, SAF-FRET, SAF-FCS, and SAF-Raman

English title Surface induced Oligomerisation and Fibrillization of alpha-Synuclein investigated by 3D-SAF, SAF-FRET, SAF-FCS, and SAF-Raman
Applicant Seeger Stefan
Number 163190
Funding scheme Project funding (Div. I-III)
Research institution Institut für Chemie Universität Zürich
Institution of higher education University of Zurich - ZH
Main discipline Physical Chemistry
Start/End 01.08.2016 - 30.04.2019
Approved amount 136'639.00
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Keywords (8)

a-Synuclein; Parkinson; neurodegenerative Krankheiten; Zellmembranen; Proteine; Proteinaggregation; Fluoreszenzspektroskopie; Raman-Spektroskopie

Lay Summary (German)

Lead
Oberflächeninduzierte Oligomerisation und Fibrillisation von alpha-Synuclein studiert mit 3D-SAF, SAF-FRET, SAF-FCS, and SAF-Raman-Mikroskopie
Lay summary

Die Alterskrankheit Parkinson wird ein immer deuticher auftretendes Phänomen in unserer Gesellschaft. Sie wird durch das Absterben von neuronalen Zellen in der “Substantia nigra” im Gehirn verursacht. Man hat sogenannte „Lewy-Bodies“ gefunden, das sind kleine Partikel, die wesentlich aus einem aggregierten Protein, a-Synuclein, bestehen. Deshalb geht man davon aus, dass a-Synuclein massgeblich an der Entstehung der Parkinson-Krankheit beteiligt ist. a-Synuclein ist ein membranaktives Protein und hat in Bezug auf chemische Signalübertragung durch Membranen eine wichtige physiologische Funktion.

 

Inhalt und Ziel des Forschungsprojekts

In der Vergangenheit haben wir durch die Entwicklung oberflächensensitiver Analysemethoden wesentliche Beiträge zum Verhalten des a-Synucleins leisten können. In dem vorliegenden Projekt werden wir das Methodenspektrum erweitern. Wir werden eine neue Technologie entwickeln, die “SAF-RAMAN-Mikroskopie”, die zum ersten Mal die Strukturen der a-Synuclein-Aggregate darstellen soll, und zwar im Rahmen der dynamisch ablaufenden Wechselwirkungsprozesse. Dieser Ansatz soll uns erlauben, Einblicke in die Faltungsstruktur des adsorbierten und aggregierten Proteins zu erhalten und darüber hinaus den Membranperforationsprozess zu studieren.

 

Wissenschaftlicher und gesellschaftlicher Kontext des Forschungsprojekts

Unsere Arbeit wird neue und wichtige Informationen über das Verhalten des Proteins a-Synuclein auf der Oberfläche von Lipiddoppelschichten ergeben. Die Ergebnisse werden ein besseres Verständnis der Wechselwirkung des Proteins auf Zelloberflächen ermöglichen und Einblick in die Aggregationsmechanismen des Proteins erlauben. Somit sollen die Arbeiten einen Beitrag zur Entwicklung von Strategien leisten, die Parkinson-Krankheit erfolgreich behandeln bzw. deren Entstehung verhindern zu können.

Direct link to Lay Summary Last update: 14.10.2015

Responsible applicant and co-applicants

Employees

Publications

Collaboration

Group / person Country
Types of collaboration
Prof. Dr. J. Robinson, Institut für Chemie, Universität Zürich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Prof. Dr. R. Riek, Lab. für Physikalische Chemie, ETH Zürich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Prof. Dr. Th. J. Schmidt, Paul Scherrer Institut Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Prof. Dr. C. Seidel, Universität Düsseldorf Germany (Europe)
- in-depth/constructive exchanges on approaches, methods or results
Prof. Dr. G. Patzke, Institut für Chemie, Universität Zürich Switzerland (Europe)
- in-depth/constructive exchanges on approaches, methods or results

Associated projects

Number Title Start Funding scheme
140402 A novel paraboloid optics for studies of single protein molecules adsorbing to supramolecular architectures 01.05.2013 Project funding (Div. I-III)
182977 Supercritical Angle Coherent Raman Spectro (SAcoR)-Imaging Facility 01.12.2018 R'EQUIP

Abstract

The age-related neurodegenerative Parkinson’s disease (PD) is becoming more and more common in our ageing society. It is caused by the apoptosis of neuronal cells especially in the substantia nigra and the locus coeruleus. In the brain of patients suffering from PD inclusion bodies, so called Lewy bodies, are found. Their main component is aggregated protein a-Synuclein, which is hence defined as the important factor for the onset of the disease. a-Synuclein is a membrane active protein, participating in membrane fusion and annealing, and is hence important for chemical signaling. The mechanism and especially the trigger of the malfunction of a-Synuclein, which is connected to a disruption of the membrane, are not fully understood.In the predecessor project we have developed different technologies based on Supercritical Angle Fluorescence (SAF) Microscopy, namely 3D-SAF, SAF-FRET, and SAF-FCS. All these technologies have been shown to be ideal tools for investigation of protein-surface interactions. In this project these technologies will be further used to study the interaction of a-Synuclein on supported lipid bilayers (SLB) which are widely accepted model systems for native cellular membranes. With this technologies we will investigate the oligomerization and fibrillization of a-synuclein on SLB surfaces. We are in particular interested in the influence of the membrane composition (in particular the charged component 1,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS) and cholesterol) and further in a more detailed investigation of the two different growth mechanism of a-Synuclein oligomers and fibrils at a SLB surface, i.e. lateral and perpendicular to the SLB plane, which we discovered recently.In addition to the methods mentioned we plan to develop a further technique, SAF-RAMAN microscopy which will enable us for the first time to record the structure of a-Synuclein aggregates and prove helix and/or ß-sheet structures from different detection volumes. This extension will give us results concerning the structure and dynamics of the aggregates and aggregation, respectively. Further, we plan to investigate with SAF-RAMAN and SAF-FRET the membrane disruption process which most likely is the molecular key process for PD.In the final part of the process we will investigate polar and total lipid extracts of porcine brain as SLB material. Here, we will come even closer to the physiological situation. Hence, we are able to answer the question if a-Synuclein interacts with lipid components or if also other membrane components are playing a role in the toxic event.
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