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Post-translational modifications of soluble α-synuclein regulate the amplification of pathological α-synuclein

Nature Neuroscience volume 26, pages 213–225 (2023)Cite this article

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Abstract

Cell-to-cell transmission and subsequent amplification of pathological proteins promote neurodegenerative disease progression. Most research on this has focused on pathological protein seeds, but how their normal counterparts, which are converted to pathological forms during transmission, regulate transmission is less understood. Here we show in cultured cells that phosphorylation of soluble, nonpathological α-synuclein (α-Syn) at previously identified sites dramatically affects the amplification of pathological α-Syn, which underlies Parkinsonʼs disease and other α-synucleinopathies, in a conformation- and phosphorylation site-specific manner. We performed LC–MS/MS analyses on soluble α-Syn purified from Parkinsonʼs disease and other α-synucleinopathies, identifying many new α-Syn post-translational modifications (PTMs). In addition to phosphorylation, acetylation of soluble α-Syn also modified pathological α-Syn transmission in a site- and conformation-specific manner. Moreover, phosphorylation of soluble α-Syn could modulate the seeding properties of pathological α-Syn. Our study represents the first systematic analysis how of soluble α-Syn PTMs affect the spreading and amplification of pathological α-Syn, which may affect disease progression.

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Fig. 1: Soluble α-Syn phosphorylation modulates pathological α-Syn amplification.
Fig. 2: Soluble α-Syn phosphorylation modulates GCI-α-Syn and LB-α-Syn amplification.
Fig. 3: Systematically identify PTMs on soluble α-Syn from different α-synucleinopathies.
Fig. 4: Soluble α-Syn acetylation modulates pathological α-Syn amplification.
Fig. 5: The effect of soluble α-Syn PTMs on pathological α-Syn transmission is conformation dependent.
Fig. 6: PTMs of soluble α-Syn modify the spreading of pathological α-Syn in primary neurons.
Fig. 7: PTMs on soluble α-Syn modify the seeding properties of pathological α-Syn.

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Data availability

Peptide identification was performed in MaxQuant (1.6.1.0) using human reference database from Uniprot (reviewed canonical and isoforms; downloaded on 1 January 2018) and the search results were exported into Scaffold 4 (Proteome Software). The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE37 partner repository with the dataset identifier PXD037994. Figure 3 and Supplementary Tables 4–8 are associated with LC–MS/MS data. Source data are provided with this paper.

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Acknowledgements

We thank Quantitative Proteomics Resource Core of School of Medicine at the University of Pennsylvania and Proteomics Core at Children’s Hospital of Philadelphia (CHOP) for performing the LC–MS/MS analysis. We thank S. Seeholzer, L. Spruce, H. Ding, H. Fazelinia and H. Lee (from CHOP) for helping with LC–MS/MS experiments. We thank D. Riddle for providing primary neurons, L. Romero for helping with quantification and all the other members of the Center for Neurodegenerative Disease Research for their support. We thank Lucy Ruoxi Shi for helping with the cartoon illustration of this work. This work was supported by NIH/NINDS Udall Center under grant NS53488 (to V.M.-Y.L.), NIA under grant U19 AG062418 (Center on Alpha-Synuclein Strains in Alzheimerʼs Disease and Related Dementias) (to V.M.-Y.L.), NIH/NINDS R01-NS103873 to E.J.P. (to V.M.-Y.L.), NIH/NINDS R01-NS128964 (to C.P.), CurePSP grant 672-2020-12 (to C.P.), the Ofer Nimerovsky Family Fund (to V.M.-Y.L.), the Jeff and Anne Keefer Fund (to V.M.-Y.L.) and the MSA Coalition Seed Grant (to C.P. and V.M.-Y.L.). The funders had no role in study design, data collection and analysis, decision to publish or preparation of the manuscript.

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Author notes

  1. These authors contributed equally: Shujing Zhang, Ruowei Zhu.

Authors and Affiliations

  1. Department of Neurology, David Geffen School of Medicine, University of California—Los Angeles, Los Angeles, CA, USA

    Shujing Zhang, Ruowei Zhu, Yuanxi Li, Luyan Zhang, Jasmine Zhang, Wenxuan Xiang, Eliot Masahiro Kagan & Chao Peng

  2. Department of Chemistry, School of Arts and Sciences, University of Pennsylvania, Philadelphia, PA, USA

    Buyan Pan & E. James Petersson

  3. The Department of Pathology and Laboratory Medicine, Institute on Aging and Center for Neurodegenerative Disease Research, the Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA

    Hong Xu, Modupe F. Olufemi, Ronald J. Gathagan, Soo-Jung Kim, John Q. Trojanowski & Virginia M.-Y. Lee

  4. Institute for Cognitive Neurodynamics, East China University of Science and Technology, Shanghai, China

    Yuanxi Li

  5. School of Mathematics, East China University of Science and Technology, Shanghai, China

    Yuanxi Li

  6. Innovent Biologics, Inc., Suzhou, China

    Xingjun Cao

  7. Alexion Pharmaceuticals, Inc., New Haven, CT, USA

    Chaoxing Yuan

  8. Section of Neuropathology, Department of Pathology and Laboratory Medicine, Ronald Reagan UCLA Medical Center and David Geffen School of Medicine, University of California—Los Angeles, Los Angeles, CA, USA

    Christopher K. Williams & Shino Magaki

  9. Section of Neuropathology, Department of Pathology and Laboratory Medicine, Department of Neurology, and Brain Research Institute, Ronald Reagan UCLA Medical Center and David Geffen School of Medicine, University of California – Los Angeles, Los Angeles, CA, USA

    Harry V. Vinters

  10. Laboratory of Molecular and Chemical Biology of Neurodegeneration, Brain Mind Institute, Swiss Federal Institute of Technology, Lausanne, Switzerland

    Hilal A. Lashuel

  11. Department of Biochemistry and Molecular Biophysics, Washington University St Louis, St Louis, MO, USA

    Benjamin A. Garcia

  12. Molecular Biology Institute, University of California, Los Angeles, Los Angeles, CA, USA

    Chao Peng

  13. Brain Research Institute, University of California, Los Angeles, Los Angeles, CA, USA

    Chao Peng

  14. Mary S. Easton Center for Alzheimer’s Research, University of California, Los Angeles, Los Angeles, CA, USA

    Chao Peng

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Contributions

C.P., V.L., S.Z. and R.Z. conceived experiments. C.P., S.Z. and R.Z. performed most of the experiments. B.P. and E.J.P. generate pY39 and pS87 α-synuclein monomer. H.X., M.F.O., R.J.G., J.Z., W.X. and E.M.K. performed cell culture experiments. L.Z. and H.A.L. performed biochemical experiments. S.-J.K. generated PFF. C.K.W., S.M. and H.V. identified and prepared human brain tissue for the preparation of pathological α-synuclein. X.C., C.Y. and B.A.G. performed LC–MS/MS experiments. Y.L. analyzed the data. C.P., J.Q.T., V.M.-Y.L., E.J.P. and H.A.L. wrote the manuscript.

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Correspondence to Virginia M.-Y. Lee or Chao Peng.

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Zhang, S., Zhu, R., Pan, B. et al. Post-translational modifications of soluble α-synuclein regulate the amplification of pathological α-synuclein. Nat Neurosci 26, 213–225 (2023). https://doi.org/10.1038/s41593-022-01239-7

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