Research Data Leeds Repository

Substitution of Met-38 to Ile in gamma-synuclein found in two patients with amyotrophic lateral sclerosis induces aggregation into amyloid

Aubrey, Liam D and Ulamec, Sabine M. and Ninkina, Natalia and Abramycheva, Natalia Y. and Limorenko, Galina and Vasili, Eftychia and Devine, Oliver M. and Wilkinson, Martin and Walko, Martin and Outeiro, Tiago F. and Chaprov, Kirill D. and Muwanga, Sarah and Amadio, Leonardo and Peters, Owen M. and Illarioshkin, Sergey N. and Ranson, Neil A and Brockwell, David J. and Buchman, Vladimir L. and Radford, Sheena E. (2023) Substitution of Met-38 to Ile in gamma-synuclein found in two patients with amyotrophic lateral sclerosis induces aggregation into amyloid. University of Leeds. [Dataset] https://doi.org/10.5518/1315

Dataset description

α-, β-, and γ-Synuclein are intrinsically disordered proteins implicated in physiological processes in the nervous system of vertebrates. α-synuclein (αSyn) is the amyloidogenic protein associated with Parkinson’s disease and certain other neurodegenerative disorders. Intensive research has focused on the mechanisms that cause αSyn to form amyloid structures, identifying its NAC region as being necessary and sufficient for amyloid assembly. Recent work has shown that a 7-residue sequence (P1) is necessary for αSyn amyloid formation. Although γ-synuclein (γSyn) is 55% identical in sequence to αSyn and its pathological deposits are also observed in association with neurodegenerative conditions, γSyn is resilient to amyloid formation in vitro. Here, we report a rare single nucleotide polymorphism (SNP) in the SNCG gene encoding γSyn, found in two patients with amyotrophic lateral sclerosis (ALS). The SNP results in the substitution of Met38 with Ile in the P1 region of the protein. These individuals also had a second, common and nonpathological, SNP in SNCG resulting in the substitution of Glu110 with Val. In vitro studies demonstrate that the Ile38 variant accelerates amyloid fibril assembly. Contrastingly, Val110 retards fibril assembly and mitigates the effect of Ile38. Substitution of residue 38 with Leu had little effect, while Val retards, and Ala increases the rate of amyloid formation. Ile38 γSyn also results in the formation of γSyn-containing inclusions in cells. The results show how a single point substitution can enhance amyloid formation of γSyn and highlight the P1 region in driving amyloid formation in another synuclein family member.

Keywords:	gamma-synuclein, ALS, aggregation, amyloid, oligomers
Subjects:	C000 - Biological sciences > C700 - Molecular biology, biophysics & biochemistry > C770 - Biophysical science C000 - Biological sciences > C400 - Genetics > C450 - Genomics
Related resources:	Location Type https://doi.org/10.1073/pnas.2309700120 Publication
License:	Creative Commons Attribution 4.0 International (CC BY 4.0)
Date deposited:	05 Jan 2024 10:47
URI:	http://archive.researchdata.leeds.ac.uk/id/eprint/1214
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 Aubrey, Liam D https://orcid.org/0000-0003-0420-8274 University of Leeds https://ror.org/024mrxd33 ror Ulamec, Sabine M. https://orcid.org/0000-0002-9415-753X University of Leeds https://ror.org/024mrxd33 ror Ninkina, Natalia https://orcid.org/0000-0001-8570-5648 Belgorod National Research University https://ror.org/044cm3z84 ror Abramycheva, Natalia Y. Research Center of Neurology, Moscow https://ror.org/05b74sw86 ror Limorenko, Galina https://orcid.org/0000-0002-6319-0257 Cardiff University https://ror.org/03kk7td41 ror École Polytechnique https://ror.org/05hy3tk52 ror Vasili, Eftychia https://orcid.org/0000-0001-9150-0104 École Polytechnique https://ror.org/05hy3tk52 ror Devine, Oliver M. University of Leeds https://ror.org/024mrxd33 ror Wilkinson, Martin https://orcid.org/0000-0001-5490-613X University of Leeds https://ror.org/024mrxd33 ror Walko, Martin https://orcid.org/0000-0002-7160-6136 University of Leeds https://ror.org/024mrxd33 ror Outeiro, Tiago F. https://orcid.org/0000-0003-1679-1727 Universitätsmedizin Göttingen https://ror.org/021ft0n22 ror Max Planck Institute for Multidisciplinary Sciences https://ror.org/03av75f26 ror Chaprov, Kirill D. https://orcid.org/0000-0002-0258-1879 Belgorod National Research University https://ror.org/044cm3z84 ror Muwanga, Sarah Cardiff University https://ror.org/03kk7td41 ror Amadio, Leonardo https://orcid.org/0000-0002-6270-536X Cardiff University https://ror.org/03kk7td41 ror Peters, Owen M. https://orcid.org/0000-0002-6824-0663 Cardiff University https://ror.org/03kk7td41 ror Illarioshkin, Sergey N. https://orcid.org/0000-0002-2704-6282 Research Center of Neurology, Moscow https://ror.org/05b74sw86 ror Ranson, Neil A https://orcid.org/0000-0002-3640-5275 University of Leeds https://ror.org/024mrxd33 ror Brockwell, David J. https://orcid.org/0000-0002-0802-5937 University of Leeds https://ror.org/024mrxd33 ror Buchman, Vladimir L. https://orcid.org/0000-0002-7631-8352 Cardiff University https://ror.org/03kk7td41 ror Belgorod National Research University https://ror.org/044cm3z84 ror Radford, Sheena E. https://orcid.org/0000-0002-3079-8039 University of Leeds https://ror.org/024mrxd33 ror Type of data: Dataset Research funders: Wellcome Trust, Royal Society, Medical Research Council, EPSRC, BBSRC Grant numbers: Wellcome 204963, Wellcome 221524/Z/20/Z, RSRP/R1/211057, Wellcome (215062/Z/18/Z), Medical Research Council UK (MR/T011149/1), Wellcome (094232), EPSRC (EP/N013573/1), BBSRC (BB/V003577/1), BBSRC (BB/T011157/1), EXC 2067/1- 390729940, SFB1286 (B8), RF (agreement No. 075-15-2021-1346), Medical Research Council (MC/PC/16030/2) Publication date: 2023 Resource language: English Publisher: University of Leeds Contact Email Address: fbslau@leeds.ac.uk Last Modified: 05 Jan 2024 23:35

Files

Documentation

readme [5kB] readme [5kB] readme [5kB] Visible to: Anyone File name: readme.txt Content type: Documentation Mime-Type: text/plain License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 5kB

Data

CD data [63kB] CD data [63kB] CD data [63kB] Visible to: Anyone File name: CD_data.xlsx Content type: Data Description: circular dichroism data Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 63kB

Differential pelleting [25kB] Differential pelleting [25kB] Differential pelleting [25kB] Visible to: Anyone File name: Differential_pelleting.xlsx Content type: Data Description: densitometry analysis of SDS-PAGE gels following differential pelleting assay Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 25kB

Liposome tht data [267kB] Liposome tht data [267kB] Liposome tht data [267kB] Visible to: Anyone File name: Liposome_ThT_data.xlsx Content type: Data Description: ThT data from liposome induced fibrillation assay Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 267kB

HPLC data [434kB] HPLC data [434kB] HPLC data [434kB] Visible to: Anyone File name: Pecentage_soluble_HPLC.xlsx Content type: Data Description: HPLC data analysis following differential pelleting assay of peptides Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 434kB

Beads and Shaking ThTs [696kB] Beads and Shaking ThTs [696kB] Beads and Shaking ThTs [696kB] Visible to: Anyone File name: Beads_and_Shaking_ThTs.zip Content type: Data Description: ThT data from beads and shaking experiments Mime-Type: application/zip License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 696kB

percent of transfected neuroblastoma cells [83kB] percent of transfected neuroblastoma cells [83kB] percent of transfected neuroblastoma cells [83kB] Visible to: Anyone File name: percent_of_transfected_neuroblastoma_cells_with_aggregates.pzfx Content type: Data Description: percent of transfected neuroblastoma cells License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 83kB

Seeded_after_beads_and_shaking_ThT [203kB] Seeded_after_beads_and_shaking_ThT [203kB] Seeded_after_beads_and_shaking_ThT [203kB] Visible to: Anyone File name: Seeded_after_beads_and_shaking_ThT.xlsx Content type: Data Description: ThT data from the experiment with beads and shaking followed by sonication and incubation under quiescent conditions. Mime-Type: application/vnd.openxmlformats-officedocument.spreadsheetml.sheet License: Creative Commons Attribution 4.0 International (CC BY 4.0) File size: 203kB