title: The novel Parkinson's disease linked mutation G51D attenuates in vitro aggregation and membrane binding of α-synuclein, and enhances its secretion and nuclear localization in cells.
abstract: A novel mutation in the α-Synuclein (α-Syn) gene "G51D" was recently identified in two familial cases exhibiting features of Parkinson's disease (PD) and multiple system atrophy (MSA). In this study, we explored the impact of this novel mutation on the aggregation, cellular and biophysical properties of α-Syn, in an attempt to unravel how this mutant contributes to PD/MSA. Our results show that the G51D mutation significantly attenuates α-Syn aggregation in vitro. Moreover, it disrupts local helix formation in the presence of SDS, decreases binding to lipid vesicles C-terminal to the site of mutation and severely inhibits helical folding in the presence of acidic vesicles. When expressed in yeast, α-Syn(G51D) behaves similarly to α-Syn(A30P), as both exhibit impaired membrane association, form few inclusions and are non-toxic. In contrast, enhanced secreted and nuclear levels of the G51D mutant were observed in mammalian cells, as well as in primary neurons, where α-Syn(G51D) was enriched in the nuclear compartment, was hyper-phosphorylated at S129 and exacerbated α-Syn-induced mitochondrial fragmentation. Finally, post-mortem human brain tissues of α-Syn(G51D) cases were examined, and revealed only partial colocalization with nuclear membrane markers, probably due to post-mortem tissue delay and fixation. These findings suggest that the PD-linked mutations may cause neurodegeneration via different mechanisms, some of which may be independent of α-Syn aggregation.
authors.affiliation: Laboratory of Molecular and Chemical Biology of Neurodegeneration, Ecole Polytechnique Fédérale de Lausanne, Station 19, CH-1015 Lausanne, Switzerland.
authors.forename: Mohamed-Bilal
authors.initials: MB
authors.lastname: Fares
authors.affiliation: Laboratory of Molecular and Chemical Biology of Neurodegeneration, Ecole Polytechnique Fédérale de Lausanne, Station 19, CH-1015 Lausanne, Switzerland.
authors.forename: Nadine
authors.initials: N
authors.lastname: Ait-Bouziad
authors.affiliation: Department of Biochemistry and Program in Structural Biology, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Igor
authors.initials: I
authors.lastname: Dikiy
authors.affiliation: Laboratory of Molecular and Chemical Biology of Neurodegeneration, Ecole Polytechnique Fédérale de Lausanne, Station 19, CH-1015 Lausanne, Switzerland.
authors.forename: Martial K
authors.initials: MK
authors.lastname: Mbefo
authors.affiliation: Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Ana
authors.initials: A
authors.lastname: Jovičić
authors.affiliation: Queen Square Brain Bank, UCL Institute of Neurology, London, UK.
authors.forename: Aoife
authors.initials: A
authors.lastname: Kiely
authors.affiliation: Queen Square Brain Bank, UCL Institute of Neurology, London, UK.
authors.forename: Janice L
authors.initials: JL
authors.lastname: Holton
authors.affiliation: Department of Biomedical Science and Technology, Konkuk University, Seoul, South Korea.
authors.forename: Seung-Jae
authors.initials: SJ
authors.lastname: Lee
authors.affiliation: Department of Genetics, Stanford University School of Medicine, Stanford, CA 94305, USA.
authors.forename: Aaron D
authors.initials: AD
authors.lastname: Gitler
authors.affiliation: Department of Biochemistry and Program in Structural Biology, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: David
authors.initials: D
authors.lastname: Eliezer
authors.affiliation: Laboratory of Molecular and Chemical Biology of Neurodegeneration, Ecole Polytechnique Fédérale de Lausanne, Station 19, CH-1015 Lausanne, Switzerland hilal.lashuel@epfl.ch.
authors.forename: Hilal A
authors.initials: HA
authors.lastname: Lashuel
mesh_terms: D001921:Brain / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D002021:Buffers; D002454:Cell Differentiation / Q000187:drug effects; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism*; D002478:Cells, Cultured; D006801:Humans; D002479:Inclusion Bodies / Q000187:drug effects / Q000378:metabolism; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D009447:Neuroblastoma / Q000473:pathology; D009474:Neurons / Q000187:drug effects / Q000378:metabolism / Q000473:pathology; D009685:Nuclear Envelope / Q000187:drug effects / Q000378:metabolism; D010300:Parkinson Disease / Q000235:genetics* / Q000473:pathology; D010766:Phosphorylation / Q000187:drug effects; D066329:Protein Aggregates / Q000187:drug effects; D066263:Protein Aggregation, Pathological / Q000235:genetics*; D011485:Protein Binding / Q000187:drug effects; D017433:Protein Structure, Secondary; D021381:Protein Transport / Q000187:drug effects; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D013347:Subcellular Fractions / Q000187:drug effects / Q000378:metabolism; D053835:Unilamellar Liposomes / Q000378:metabolism; D051844:alpha-Synuclein / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D002021:Buffers; D066329:Protein Aggregates; D053835:Unilamellar Liposomes; D051844:alpha-Synuclein; D012967:Sodium Dodecyl Sulfate
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