N‐terminal region of α‐synuclein is essential for the fatty acid‐induced oligomerization of the molecules

Karube, Hiroki; Sakamoto, Masahiro; Arawaka, Shigeki; Hara, Shotaro; Sato, Hiroyasu; Ren, Chao; Goto, Saori; Koyama, Shingo; Wada, Manabu; Kawanami, Toru; Kurita, Keiji; Kato, Takeshi

doi:10.1016/j.febslet.2008.10.001

Cited by 52 publications

(49 citation statements)

References 35 publications

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“…We used C-terminal tags where ␣-synuclein is connected to the tag via a linker. The N-terminal region needs to remain tag-free for ␣-synuclein to be toxic, presumably because it is essential for membrane interactions and oligomerization (52). The C terminus is highly acidic and tends to inhibit protein aggregation due to other protein-protein interactions (53)(54)(55).…”

Section: Discussionmentioning

confidence: 99%

Aggregate Clearance of α-Synuclein in Saccharomyces cerevisiae Depends More on Autophagosome and Vacuole Function Than on the Proteasome

Petroi

Popova

Taheri-Talesh

et al. 2012

Journal of Biological Chemistry

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Background:The hallmark of Parkinson disease is ␣-synuclein aggregation, whose cytotoxic effects are reproducible in yeast. Results: ␣-Synuclein aggregate clearance is impaired by vacuolar protease inhibitor PMSF but not by proteasome inhibitor MG132. Conclusion: Yeast recovers from ␣-synuclein-induced toxicity by clearing aggregates via autophagy and vacuolar pathways. Significance: The data provide insight into the mechanisms used by yeast cells to clear toxic aggregated proteins.

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Section: Discussionmentioning

confidence: 99%

Aggregate Clearance of α-Synuclein in Saccharomyces cerevisiae Depends More on Autophagosome and Vacuole Function Than on the Proteasome

Petroi

Popova

Taheri-Talesh

et al. 2012

Journal of Biological Chemistry

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“…Recombinant non-phosphorylated a-Syn proteins were purified from Escherichia coli as described previously (14). Phosphorylated a-Syn proteins were made by incubation of 100 g of non-phosphorylated proteins in reaction buffer (20 mM Tris-HCl, pH 7.5, 50 mM KCl, 10 mM MgCl 2 , 200 M ATP) containing 1,000 units of recombinant CK2 protein (New England Biolabs, Beverly, MA) for 16 h at 37°C.…”

Section: Sporadic Parkinson Disease (Spd)mentioning

confidence: 99%

Phosphorylated α-Synuclein at Ser-129 Is Targeted to the Proteasome Pathway in a Ubiquitin-independent Manner

Machiya¹,

Hara²,

Arawaka

et al. 2010

Journal of Biological Chemistry

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106

115

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␣-Synuclein (a-Syn) is a major component of fibrillar aggregates in Lewy bodies (LBs), a characteristic hallmark of Parkinson disease. Almost 90% of a-Syn deposited in LBs is phosphorylated at Ser-129. However, the role of Ser-129-phosphorylated a-Syn in the biogenesis of LBs remains unclear. Here, we investigated the metabolism of Ser-129-phosphorylated a-Syn. In SH-SY5Y cells, inhibition of protein phosphatase 2A/1 by okadaic acid, and inhibition of the proteasome pathway by MG132 or lactacystin accumulated Ser-129-phosphorylated a-Syn. However, these inhibitions did not alter the amounts of total a-Syn within the observation time. Inhibition of the autophagy-lysosome pathway by 3-methyladenine or chloroquine accumulated Ser-129-phosphorylated a-Syn in parallel to total a-Syn during longer incubations. Experiments using cycloheximide showed that Ser-129-phosphorylated a-Syn diminished rapidly (t1 ⁄ 2 ‫؍‬ 54.9 ؎ 6.4 min), in contrast to the stably expressed total a-Syn. The short half-life of Ser-129-phosphorylated a-Syn was blocked by MG132 to a greater extent than okadaic acid. In rat primary cortical neurons, either MG132, lactacystin, or okadaic acid accumulated Ser-129-phosphorylated a-Syn. Additionally, we did not find that phosphorylated a-Syn was ubiquitinated in the presence of proteasome inhibitors. These data show that Ser-129-phosphorylated a-Syn is targeted to the proteasome pathway in a ubiquitin-independent manner, in addition to undergoing dephosphorylation. The proteasome pathway may play a role in the biogenesis of Ser-129-phosphorylated a-Syn-rich LBs. Sporadic Parkinson disease (sPD)3 is characterized pathologically by a loss of dopaminergic neurons in the substantia nigra pars compacta and the presence of intracytoplasmic inclusions called Lewy bodies (LBs) and Lewy neurites (LNs) in surviving neurons. ␣-Synuclein (a-Syn) is a major component of fibrillar aggregates in LBs and LNs. Accumulating lines of evidence have shown that prefibrillar intermediates of a-Syn, such as soluble oligomers or protofibrils, play a toxic role in degeneration of dopaminergic neurons, and mature fibrils of a-Syn contribute toward this toxicity to a lesser extent (1-4). Therefore, the process of a-Syn aggregation eventually forming LBs is proposed to play a causative role in neuronal degeneration of PD (5, 6). Immunohistochemical and biochemical studies have revealed that ϳ90% of a-Syn deposited in LBs is phosphorylated at serine 129 (Ser-129) (7,8). In contrast, the portion of phosphorylated a-Syn in normal brains is known to be only about 4% (7) or less than the limits of quantification of the assays used (8). This discrepancy implicates a pathogenic role of Ser-129-phosphorylated a-Syn in the biogenesis of LBs (7, 9). One possibility is that the Ser-129-phosphorylation promotes the aggregation-prone property of a-Syn. To elucidate this issue, several in vitro studies have been performed. However, the accelerating effect of phosphorylation on fibril formation of a-Syn is controversial at present (7, 10). Anoth...

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“…This hypothesis has been constructed and tested largely using in vitro and in vivo studies on AD and polyglutamine expansion diseases [10,29,30]. In the case of PD, soluble oligomers of alpha-synuclein can be induced by adding polyunsaturated fatty acids to cultured cells [79][80][81], and fatty acid-associated soluble oligomers of alpha-synuclein are observed in alphasynuclein transgenic mice brains and PD brains [79]. However, to detect fatty acid-associated soluble oligomers of alpha-synuclein, it is necessary to treat cell lysates and brain homogenates with a lipid removal procedure, such as heat or methanol/chloroform [79][80][81].…”

Section: Discussionmentioning

confidence: 98%

Heat Shock Proteins as Suppressors of Accumulation of Toxic Prefibrillar Intermediates and Misfolded Proteins in Neurodegenerative Diseases

Arawaka¹,

Machiya²,

Kato

2010

CPB

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The most characteristic feature of neurodegenerative diseases, including Alzheimer's disease, Parkinson's disease, amyotrophic lateral sclerosis and Huntington's disease, is the occurrence of extra- or intracellular fibrillar aggregates containing misfolded proteins with beta-sheet conformation. These aggregates are composed of distinct proteins in each neurodegenerative disease. However, mutations in genes encoding major constituents of aggregates, such as Abeta, tau, alpha-synuclein, SOD1 and huntingtin, have been identified to causally associate with familial forms of the diseases. Biochemical studies demonstrate that these mutant and some wild-type proteins tend to be misfolded or form aggregates. It has been proposed that these diseases are caused by a common mechanism involving misfolded proteins that trigger a toxic cascade leading to neuronal degeneration. This hypothesis is the basis of the therapeutic potential of heat shock proteins (HSPs), which prevent protein misfolding and aggregation. Transgenic animal models of the diseases have demonstrated that induction or overexpression of HSPs can suppress neuronal dysfunction and degeneration. Do the results promise clinical success for HSP-based therapy in neurodegenerative diseases? Recent findings regarding the pathogenic species generated during fibril formation have highlighted some of the beneficial and problematic aspects of HSP-based therapy. In this review, we focus on the pathogenic role of prefibrillar intermediates, including soluble oligomers and protofibrils, on neurodegeneration, and the relationship between prefibrillar intermediates and the proteins targeted by HSPs. We discuss in vitro and in vivo experimental data showing that HSPs counteract disease progression by acting as suppressors of toxic prefibrillar intermediates and toxic misfolded proteins in neurodegenerative diseases.

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N‐terminal region of α‐synuclein is essential for the fatty acid‐induced oligomerization of the molecules

Cited by 52 publications

References 35 publications

Aggregate Clearance of α-Synuclein in Saccharomyces cerevisiae Depends More on Autophagosome and Vacuole Function Than on the Proteasome

Aggregate Clearance of α-Synuclein in Saccharomyces cerevisiae Depends More on Autophagosome and Vacuole Function Than on the Proteasome

Phosphorylated α-Synuclein at Ser-129 Is Targeted to the Proteasome Pathway in a Ubiquitin-independent Manner

Heat Shock Proteins as Suppressors of Accumulation of Toxic Prefibrillar Intermediates and Misfolded Proteins in Neurodegenerative Diseases

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