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Studying α-Synuclein Conformation by Intact-Cell Cross-Linking

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Alpha-Synuclein

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1948))

Abstract

β-Sheet-rich aggregates of α-synuclein (αS) are the hallmark neuropathology of Parkinson’s disease (PD) and related synucleinopathies, whereas the native conformations of αS in healthy cells are under debate. Cross-linking analyses in intact cells detect a large portion of endogenous αS in apparent multimeric states, most notably as putative tetramers (αS60) that run around 60 kDa on SDS-PAGE, but also point at the dynamic nature of cellular αS states. Standardization of αS cross-linking methods will facilitate efforts to study the effects of genetic, pharmacological, and environmental factors on αS conformation. Here, we present detailed protocols for cross-linking cellular αS multimers in cultured cells and brain tissues. These protocols will benefit future studies aimed at characterizing αS conformation in its cellular environment, both at steady state and upon perturbation, be it chronic or acute.

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Authors and Affiliations

  1. Ann Romney Center for Neurologic Diseases, Brigham and Women’s Hospital and Harvard Medical School, Boston, MA, USA

    Thibaut Imberdis, Saranna Fanning, Andrew Newman, Nagendran Ramalingam & Ulf Dettmer

Authors
  1. Thibaut Imberdis
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  2. Saranna Fanning
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  3. Andrew Newman
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  4. Nagendran Ramalingam
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  5. Ulf Dettmer
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Corresponding author

Correspondence to Ulf Dettmer .

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Editors and Affiliations

  1. Dementia Research Institute, University College London, London, UK

    Tim Bartels

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Imberdis, T., Fanning, S., Newman, A., Ramalingam, N., Dettmer, U. (2019). Studying α-Synuclein Conformation by Intact-Cell Cross-Linking. In: Bartels, T. (eds) Alpha-Synuclein. Methods in Molecular Biology, vol 1948. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9124-2_8

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  • DOI: https://doi.org/10.1007/978-1-4939-9124-2_8

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9123-5

  • Online ISBN: 978-1-4939-9124-2

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