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Published on: June 27, 2018
Huntingtin cleavage induced by thrombin in vitro
Fang Lin1, Junchao Wu, Yan Wang
1Department of Pharmacology, Laboratory of Aging and Nervous Diseases, Soochow University School of Medicine, Suzhou 215123, China.
Acta Biochimica Et Biophysica Sinica
|January 11, 2007
Summary
This study shows that thrombin cleaves Huntingtin (Htt) protein in vitro. This cleavage, occurring at amino acid 183, produces an 80 kDa fragment, suggesting a new pathway in Huntington
Area of Science:
- Biochemistry
- Molecular Biology
- Neuroscience
Background:
- Huntington's disease is caused by mutations in the Huntingtin (Htt) gene.
- Sequence analysis suggested a potential thrombin cleavage site in the N-terminal region of Htt.
Purpose of the Study:
- To investigate whether thrombin can cleave the Huntingtin protein.
- To identify the specific cleavage site and resulting fragments.
Main Methods:
- Expressed the N-terminal fragment (1-969) of wild-type Htt in MCF-7 cells.
- Incubated cell lysates with varying concentrations of thrombin in vitro.
- Analyzed Htt fragments using SDS-PAGE and Western blotting.
- Utilized hirudin, a thrombin inhibitor, to confirm specificity.
Main Results:
- Thrombin cleaved Htt (1-969) to produce an approximately 80 kDa fragment.
- Cleavage occurred at amino acid 183, consistent with predictions.
- Hirudin inhibited the production of the 80 kDa fragment, confirming thrombin's role.
Conclusions:
- Provides the first evidence that thrombin cleaves Htt in vitro at amino acid 183.
- Suggests a potential in vivo role for thrombin in Htt processing that warrants further investigation.
