Preparation and characterization of stable α-synuclein lipoprotein particles

Cédric Eichmann, Silvia Campioni, Julia Kowal, Innokentiy Maslennikov, Juan Gerez, Xiaoxia Liu, Joeri Verasdonck, Nadezhda Nespovitaya, Senyon Choe, Beat H. Meier, Paola Picotti, Jose Rizo-Rey, Henning Stahlberg, Roland Riek

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27 Scopus citations

Abstract

Multiple neurodegenerative diseases are caused by the aggregation of the human α-Synuclein (α-Syn) protein. α-Syn possesses high structural plasticity and the capability of interacting with membranes. Both features are not only essential for its physiological function but also play a role in the aggregation process. Recently it has been proposed that α-Syn is able to form lipid-protein particles reminiscent of high-density lipoproteins. Here, we present a method to obtain a stable and homogeneous population of nanometer-sized particles composed of α-Syn and anionic phospholipids. These particles are called α-Syn lipoprotein (nano) particles to indicate their relationship to high-density lipoproteins formed by human apolipoproteins in vivo and of in vitro self-assembling phospholipid bilayer nanodiscs. Structural investigations of the α-Syn lipoprotein particles by circular dichroism (CD) and magic angle solid-state nuclear magnetic resonance (MAS SS-NMR) spectroscopy establish that α-Syn adopts a helical secondary structure within these particles. Based on cryo-electron microscopy (cryo-EM) and dynamic light scattering (DLS) α-Syn lipoprotein particles have a defined size with a diameter of ∼23 nm. Chemical cross linking in combination with solution-state NMR and multiangle static light scattering (MALS) of α-Syn particles reveal a high order protein-lipid entity composed of α 8-10 α-Syn molecules. The close resemblance in size between cross-linked in vitro-derived α-Syn lipoprotein particles and a cross-linked species of endogenous α-Syn from SH-SY5Y human neuroblastoma cells indicates a potential functional relevance of α-Syn lipoprotein nanoparticles.

Original languageEnglish (US)
Pages (from-to)8516-8527
Number of pages12
JournalJournal of Biological Chemistry
Volume291
Issue number16
DOIs
Publication statusPublished - Apr 15 2016

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ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Eichmann, C., Campioni, S., Kowal, J., Maslennikov, I., Gerez, J., Liu, X., ... Riek, R. (2016). Preparation and characterization of stable α-synuclein lipoprotein particles. Journal of Biological Chemistry, 291(16), 8516-8527. https://doi.org/10.1074/jbc.M115.707968