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Phosphorylated alpha-synuclein is ubiquitinated in alpha-synucleinopathy lesions.
Masato Hasegawa1, Hideo Fujiwara, Takashi Nonaka
1Department of Molecular Neurobiology, Tokyo Institute of Psychiatry, Tokyo Metropolitan Organization for Medical Research, 2-1-8 Kamikitazawa, Setagaya-ku, Japan. masato@prit.go.jp
The Journal of Biological Chemistry
|October 16, 2002
Summary
Phosphorylated alpha-synuclein in neurodegenerative diseases is ubiquitinated, forming higher molecular mass species. This suggests ubiquitination targets phosphorylated alpha-synuclein in alpha-synucleinopathies.
Area of Science:
- Neurodegenerative disease research
- Protein biochemistry
- Molecular neuroscience
Background:
- Alpha-synuclein aggregates in alpha-synucleinopathies like Parkinson's disease.
- Previously, phosphorylated alpha-synuclein at Ser-129 was identified in synucleinopathy brains.
Purpose of the Study:
- Investigate higher molecular mass species of phosphorylated alpha-synuclein.
- Determine the biochemical characteristics and potential modifications of these species.
Main Methods:
- Analysis of Sarkosyl-insoluble fractions from synucleinopathy brains.
- Immunoblotting with anti-ubiquitin and anti-alpha-synuclein antibodies.
- In vitro ubiquitination assays.
- Cyanogen bromide cleavage and protein sequencing.
Main Results:
- Identified 22 and 29 kDa phosphorylated alpha-synuclein species.
- These species reacted with anti-ubiquitin antibodies and comigrated with in vitro ubiquitinated alpha-synuclein.
- Cyanogen bromide cleavage and sequencing confirmed mono- and diubiquitination of alpha-synuclein.
Conclusions:
- Phosphorylated alpha-synuclein is targeted for mono- and diubiquitination in alpha-synucleinopathy brains.
- This ubiquitination may play a role in the pathogenesis of these neurodegenerative diseases.