Oxidative stress-induced posttranslational modifications of alpha-synuclein: specific modification of alpha-synuclein by 4-hydroxy-2-nonenal increases dopaminergic toxicity.
Aggregation and neurotoxicity of misfolded alpha-synuclein (αSyn) are crucial mechanisms for progressive dopaminergic neurodegeneration associated with Parkinson's disease (PD). Posttranslational modifications (PTMs) of αSyn caused by oxidative stress, including modification by 4-hydroxy-2-nonenal (HNE-αSyn), nitration (n-αSyn), and oxidation (o-αSyn), have been implicated to promote oligomerization of αSyn. However, it is yet unclear if these PTMs lead to different types of oligomeric intermediates. Moreover, little is known about which PTM-derived αSyn species exerts toxicity to dopaminergic cells. In this study, we directly compared aggregation characteristics of HNE-αSyn, n-αSyn, and o-αSyn. Generally, all of them promoted αSyn oligomerization. Particularly, HNE-αSyn and n-αSyn were more prone to forming oligomers than unmodified αSyn. Moreover, these PTMs prevented the formation of amyloid-like fibrils, although HNE-αSyn and o-αSyn were able to generate protofibrillar structures. The cellular effects associated with distinct PTMs were studied by exposing modified αSyn to dopaminergic Lund human mesencephalic (LUHMES) neurons. The cellular toxicity of HNE-αSyn was significantly higher than other PTM species. Furthermore, we tested the toxicity of HNE-αSyn in dopaminergic LUHMES cells and other cell types with low tyrosine hydroxylase (TH) expression, and additionally analyzed the loss of TH-immunoreactive cells in HNE-αSyn-treated LUHMES cells. We observed a selective toxicity of HNE-αSyn to neurons with higher TH expression. Further mechanistic studies showed that HNE-modification apparently increased the interaction of extracellular αSyn with neurons. Moreover, exposure of differentiated LUHMES cells to HNE-αSyn triggered the production of intracellular reactive oxygen species, preceding neuronal cell death. Antioxidant treatment effectively protected cells from the damage triggered by HNE-αSyn. Our findings suggest a specific pathological effect of HNE-αSyn on dopaminergic neurons.
Institute of Biochemistry (Emil-Fischer-Center), Friedrich-Alexander-University of Erlangen Nürnberg, 91054 Erlangen, Germany. wei.xiang@biochem.uni-erlangen.de
Wei
W
Xiang
Johannes C M
JC
Schlachetzki
Stefan
S
Helling
Julia C
JC
Bussmann
Marvin
M
Berlinghof
Tilman E
TE
Schäffer
Katrin
K
Marcus
Jürgen
J
Winkler
Jochen
J
Klucken
Cord-Michael
CM
Becker
D000447:Aldehydes / Q000737:chemistry / Q000633:toxicity*; D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D003432:Cross-Linking Reagents / Q000737:chemistry / Q000633:toxicity*; D059290:Dopaminergic Neurons / Q000187:drug effects* / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology; D006801:Humans; D008636:Mesencephalon / Q000166:cytology; D018384:Oxidative Stress; D055503:Protein Multimerization; D011499:Protein Processing, Post-Translational; D017382:Reactive Oxygen Species / Q000378:metabolism; D051844:alpha-Synuclein / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*
D000447:Aldehydes; D003432:Cross-Linking Reagents; D017382:Reactive Oxygen Species; D051844:alpha-Synuclein; C027576:4-hydroxy-2-nonenal