TY - JOUR
T1 - Preparation and characterization of stable α-synuclein lipoprotein particles
AU - Eichmann, Cédric
AU - Campioni, Silvia
AU - Kowal, Julia
AU - Maslennikov, Innokentiy
AU - Gerez, Juan
AU - Liu, Xiaoxia
AU - Verasdonck, Joeri
AU - Nespovitaya, Nadezhda
AU - Choe, Senyon
AU - Meier, Beat H.
AU - Picotti, Paola
AU - Rizo-Rey, Jose
AU - Stahlberg, Henning
AU - Riek, Roland
N1 - Publisher Copyright:
© 2016 by The American Society for Biochemistry and Molecular Biology, Inc.
PY - 2016/4/15
Y1 - 2016/4/15
N2 - 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.
AB - 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.
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U2 - 10.1074/jbc.M115.707968
DO - 10.1074/jbc.M115.707968
M3 - Article
C2 - 26846854
AN - SCOPUS:84965141075
SN - 0021-9258
VL - 291
SP - 8516
EP - 8527
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 16
ER -