Systematic comparison of the effects of alpha-synuclein mutations on its oligomerization and aggregation.
Aggregation of alpha-synuclein (ASYN) in Lewy bodies and Lewy neurites is the typical pathological hallmark of Parkinson's disease (PD) and other synucleinopathies. Furthermore, mutations in the gene encoding for ASYN are associated with familial and sporadic forms of PD, suggesting this protein plays a central role in the disease. However, the precise contribution of ASYN to neuronal dysfunction and death is unclear. There is intense debate about the nature of the toxic species of ASYN and little is known about the molecular determinants of oligomerization and aggregation of ASYN in the cell. In order to clarify the effects of different mutations on the propensity of ASYN to oligomerize and aggregate, we assembled a panel of 19 ASYN variants and compared their behaviour. We found that familial mutants linked to PD (A30P, E46K, H50Q, G51D and A53T) exhibited identical propensities to oligomerize in living cells, but had distinct abilities to form inclusions. While the A30P mutant reduced the percentage of cells with inclusions, the E46K mutant had the opposite effect. Interestingly, artificial proline mutants designed to interfere with the helical structure of the N-terminal domain, showed increased propensity to form oligomeric species rather than inclusions. Moreover, lysine substitution mutants increased oligomerization and altered the pattern of aggregation. Altogether, our data shed light into the molecular effects of ASYN mutations in a cellular context, and established a common ground for the study of genetic and pharmacological modulators of the aggregation process, opening new perspectives for therapeutic intervention in PD and other synucleinopathies.
Department of NeuroDegeneration and Restorative Research, Center for Nanoscale Microscopy and Molecular Physiology of the Brain University Medical Goettingen, Goettingen, Germany.
Diana F
DF
Lázaro
Department of NeuroDegeneration and Restorative Research, Center for Nanoscale Microscopy and Molecular Physiology of the Brain University Medical Goettingen, Goettingen, Germany.
Eva F
EF
Rodrigues
Department of Neurology, Ulm University, Ulm, Germany.
Ramona
R
Langohr
Georg August University, Institute for Microbiology and Genetics Dept. of Molecular Microbiology and Genetics, Goettingen, Germany.
Hedieh
H
Shahpasandzadeh
Laboratório de Citotoxicidade e Genotoxicidade, Departamento de Bioquímica - Instituto de Química Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Thales
T
Ribeiro
Department of NeuroDegeneration and Restorative Research, Center for Nanoscale Microscopy and Molecular Physiology of the Brain University Medical Goettingen, Goettingen, Germany; Instituto de Medicina Molecular, Faculdade de Medicina da Universidade de Lisboa, Lisboa, Portugal.
Patrícia
P
Guerreiro
Department of NeuroDegeneration and Restorative Research, Center for Nanoscale Microscopy and Molecular Physiology of the Brain University Medical Goettingen, Goettingen, Germany.
Ellen
E
Gerhardt
Department of Neuro and Sensory Physiology, University of Göttingen Medical Center c/o European Neuroscience Institute Göttingen, Göttingen, Germany.
Katharina
K
Kröhnert
Department of Molecular Neurology, University Hospital Erlangen, Friedrich-Alexander University Erlangen-Nürnberg, Erlangan, Germany.
Jochen
J
Klucken
Laboratório de Citotoxicidade e Genotoxicidade, Departamento de Bioquímica - Instituto de Química Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Marcos D
MD
Pereira
Georg August University, Institute for Microbiology and Genetics Dept. of Molecular Microbiology and Genetics, Goettingen, Germany.
Blagovesta
B
Popova
Institute for Neuropathology, University Medical Center Goettingen, Goettingen, Germany.
Niels
N
Kruse
Institute for Neuropathology, University Medical Center Goettingen, Goettingen, Germany; The Department for neurosurgery at UMG and Paracelsus-Elena-Klinik, Kassel, Germany.
Brit
B
Mollenhauer
Department of Neuro and Sensory Physiology, University of Göttingen Medical Center c/o European Neuroscience Institute Göttingen, Göttingen, Germany.
Silvio O
SO
Rizzoli
Georg August University, Institute for Microbiology and Genetics Dept. of Molecular Microbiology and Genetics, Goettingen, Germany.
Gerhard H
GH
Braus
Department of Neurology, Ulm University, Ulm, Germany.
Karin M
KM
Danzer
Department of NeuroDegeneration and Restorative Research, Center for Nanoscale Microscopy and Molecular Physiology of the Brain University Medical Goettingen, Goettingen, Germany; Laboratório de Citotoxicidade e Genotoxicidade, Departamento de Bioquímica - Instituto de Química Universidade Federal do Rio de Janeiro, Rio de Janeiro, Brazil.
Tiago F
TF
Outeiro
D002460:Cell Line; D006801:Humans; D016631:Lewy Bodies / Q000378:metabolism / Q000473:pathology; D008247:Lysosomes / Q000378:metabolism / Q000473:pathology; D016297:Mutagenesis, Site-Directed; D010300:Parkinson Disease / Q000235:genetics* / Q000473:pathology; D010766:Phosphorylation; D017354:Point Mutation; D066263:Protein Aggregation, Pathological / Q000235:genetics*; D051844:alpha-Synuclein / Q000235:genetics* / Q000378:metabolism
D051844:alpha-Synuclein