Research Data Leeds Repository

Residues 2 to 7 of α-synuclein regulate amyloid formation via lipid-dependent and lipid-independent pathways

Dewison, Katherine and Radford, Sheena (2024) Residues 2 to 7 of α-synuclein regulate amyloid formation via lipid-dependent and lipid-independent pathways. University of Leeds. [Dataset] https://doi.org/10.5518/1422

Dataset description

Here, the role of residues 2-7 in the N-terminal region of αSyn are investigated in terms of their ability to regulate amyloid fibril formation in vitro and in vivo. Deletion of these residues (αSynΔN7) slows the rate of fibril formation in vitro and reduces the capacity of the protein to be recruited by wild-type (αSynWT) fibril seeds, despite cryo-EM showing a fibril structure consistent with those of full-length αSyn. Strikingly, fibril formation of αSynΔN7 is not induced by liposomes, despite the protein binding to liposomes with similar affinity to αSynWT. A Caenorhabditis elegans model also showed that αSynΔN7::YFP forms few puncta and lacks motility and lifespan defects typified by expression of αSynWT::YFP.

Subjects:	C000 - Biological sciences > C700 - Molecular biology, biophysics & biochemistry
Divisions:	Faculty of Biological Sciences > Astbury Centre for Structural Molecular Biology
Related resources:	Location Type https://doi.org/10.1101/2024.05.24.595537 Publication
License:	Creative Commons Attribution 4.0 International (CC BY 4.0)
Date deposited:	29 May 2024 20:05
URI:	https://archive.researchdata.leeds.ac.uk/id/eprint/1282
Additional details Additional details Creators: Creators ORCID Other ID Email Primary affiliation Primary affiliation ID Primary affiliation ID type Secondary affiliation Secondary affiliation ID Secondary affiliation ID type Dewison, Katherine https://orcid.org/0000-0002-7196-7687 University of Leeds https://ror.org/024mrxd33 ror Radford, Sheena https://orcid.org/0000-0002-3079-8039 University of Leeds https://ror.org/024mrxd33 ror Type of data: Dataset Research funders: Wellcome Trust, MRC, BBSRC, The Royal Society Grant numbers: 094232/Z/10/Z, 104918MA, MR/N013840/1, MR/T011149/1, BB/W007649/1, BB/T008059/1, 222373/Z/21/Z, 215062/Z/18/Z, 204963, RSRP/R1/211057 Publication date: 2024 Resource language: English Publisher: University of Leeds Contact Email Address: S.E.Radford@leeds.ac.uk Last Modified: 31 May 2024 10:03

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Documentation

README_Dewison_2024.txt [10kB] README_Dewison_2024.txt [10kB] README_Dewison_2024.txt [10kB] Visible to: Anyone File name: README_Dewison_2024.txt Content type: Documentation Mime-Type: text/plain License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 10kB

Data

Figure1 [70kB] Figure1 [70kB] Figure1 [70kB] Visible to: Anyone File name: DOI_Figure1.xlsx Content type: Data Description: Deletion of residues 2-7 of αSyn slows amyloid formation in vitro Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 70kB

Figure2 [90kB] Figure2 [90kB] Figure2 [90kB] Visible to: Anyone File name: DOI_Figure2.xlsx Content type: Data Description: Residues 2-7 of αSyn are required to elongate αSynWT fibril seeds Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 90kB

Figure3 [98kB] Figure3 [98kB] Figure3 [98kB] Visible to: Anyone File name: DOI_Figure3.xlsx Content type: Data Description: Residues 2-7 of αSyn are not necessary for DMPS liposome binding Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 98kB

Figure4 [105kB] Figure4 [105kB] Figure4 [105kB] Visible to: Anyone File name: DOI_Figure4.xlsx Content type: Data Description: Residues 2-7 of αSyn are critical for lipid-mediated fibrillation Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 105kB

Figure5 [29kB] Figure5 [29kB] Figure5 [29kB] Visible to: Anyone File name: DOI_Figure5.xlsx Content type: Data Description: C. elegans expressing αSynΔN7::YFP exhibit fewer aggregates and motility defects than those expressing αSynWT::YFP Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 29kB

FigureS1 [72kB] FigureS1 [72kB] FigureS1 [72kB] Visible to: Anyone File name: DOI_SI_Figure1.xlsx Content type: Data Description: Amyloid formation kinetics of the previously-studied deletion variants of αSyn (αSynΔP1, αSynΔP2, and αSynΔΔ) Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 72kB

FigureS4 [226kB] FigureS4 [226kB] FigureS4 [226kB] Visible to: Anyone File name: DOI_SI_Figure4.xlsx Content type: Data Description: Effect of mixing αSynWT and αSynΔN7 monomers on amyloid formation Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 226kB

FigureS6 [58kB] FigureS6 [58kB] FigureS6 [58kB] Visible to: Anyone File name: DOI_SI_Figure6.xlsx Content type: Data Description: T50 values for the seeded fibril growth assays shown in Fig. 2 Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 58kB

FigureS7 [88kB] FigureS7 [88kB] FigureS7 [88kB] Visible to: Anyone File name: DOI_SI_Figure7.xlsx Content type: Data Description: Seeded fibril growth kinetics from unsonicated fibril seeds Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 88kB

FigureS9 [62kB] FigureS9 [62kB] FigureS9 [62kB] Visible to: Anyone File name: DOI_SI_Figure9.xlsx Content type: Data Description: 1H-15N HMQC NMR spectra of αSynWT and αSynΔN7 binding to DMPS liposomes at different temperatures Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 62kB

FigureS10 [21kB] FigureS10 [21kB] FigureS10 [21kB] Visible to: Anyone File name: DOI_SI_Figure10.xlsx Content type: Data Description: Monitoring DMPS membrane fluidity and lipid packing upon incubation with αSynWT and αSynΔN7 monomers Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 21kB

FigureS12 [97kB] FigureS12 [97kB] FigureS12 [97kB] Visible to: Anyone File name: DOI_SI_Figure12.xlsx Content type: Data Description: Characterisation of the behaviour of αSynΔP1 in the presence of DMPS liposomes Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 97kB

FigureS13 [25kB] FigureS13 [25kB] FigureS13 [25kB] Visible to: Anyone File name: DOI_SI_Figure13.xlsx Content type: Data Description: Far-UV CD spectra of alanine scan variants of αSyn (residues 2-7) Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 25kB

FigureS14 [78kB] FigureS14 [78kB] FigureS14 [78kB] Visible to: Anyone File name: DOI_SI_Figure14.xlsx Content type: Data Description: Fibril formation kinetics of alanine scan variants of αSyn in the presence of DMPS liposomes Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 78kB

FigureS15 [16kB] FigureS15 [16kB] FigureS15 [16kB] Visible to: Anyone File name: DOI_SI_Figure15.xlsx Content type: Data Description: Densitometry of the western blots showing protein expression levels of αSynWT::YFP and αSynΔN7::YFP in the body wall muscle cells of C. elegans Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 16kB

FigureS16 [16kB] FigureS16 [16kB] FigureS16 [16kB] Visible to: Anyone File name: DOI_SI_Figure16.xlsx Content type: Data Description: Lifespan data for aSynDN7 C. elegans strain compared with aSynWT and N2 Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 16kB

C.elegans images [2GB] C.elegans images [2GB] C.elegans images [2GB] Visible to: Anyone File name: C_elegans_images.zip Content type: Data Description: zipped folder containing all C. elegans confocal images Mime-Type: application/zip License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 2GB

SDS-PAGE gels [144MB] SDS-PAGE gels [144MB] SDS-PAGE gels [144MB] Visible to: Anyone File name: SDS-PAGE-gels.zip Content type: Data Description: folder containing all SDS-PAGE gels used for densitometry calculations Mime-Type: application/zip License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 144MB