One-pot semisynthesis of exon 1 of the Huntingtin protein: new tools for elucidating the role of posttranslational modifications in the pathogenesis of Huntington's disease
| Autor: | Jae Sun Jeong, Francesco Simone Ruggeri, Zhe Ming Wang, Annalisa Ansaloni, Hilal A. Lashuel, Giovanni Dietler |
|---|---|
| Rok vydání: | 2013 |
| Předmět: |
huntington's disease
Huntingtin Molecular Sequence Data Nerve Tissue Proteins Catalysis Exon huntingtin exon 1 Huntington's disease medicine Huntingtin Protein Humans Amino Acid Sequence Phosphorylation chemistry.chemical_classification phosphorylation aggregation General Chemistry General Medicine Exons medicine.disease Semisynthesis Glutamine Enzyme chemistry Biochemistry huntingtin exon1 protein semisynthesis Protein Processing Post-Translational |
| Zdroj: | Angewandte Chemie-International Edition, 53(7), 1928-1933 Angewandte Chemie-International Edition 53 (2014) 7 |
| ISSN: | 1521-3773 1433-7851 |
| Popis: | The natural enzymes involved in regulating many of the posttranslational modifications (PTMs) within the first 17 residues (Nt17) of Huntingtin exon 1 (Httex1) remain unknown. A semisynthetic strategy that allows the site-specific introduction of PTMs within Nt17 by using expressed protein ligation (EPL) was developed. This strategy was used to produce untagged wild-type (wt) and T3-phosphorylated (pT3) Httex1 containing 23 glutamine residues (Httex1-23Q). Our studies show that pT3 significantly slows the oligomerization and fibrillization of Httex1-23Q and that Httex1 variants containing polyQ repeats below the pathogenic threshold readily aggregate and form fibrils in vitro. These findings suggest that crossing the polyQ pathogenic threshold is not essential for Httex1 aggregation. The ability to produce wt or site-specifically modified tag-free Httex1 should facilitate determining its structure and the role of N-terminal PTMs in regulating the functions of Htt in health and disease. Tailor-made: A one-pot semisynthetic strategy that enables the site-specific introduction of posttranslational modifications within the N terminus of exon 1 of the Huntingtin protein (Httex1) is reported. This approach was applied to generate untagged wild-type and T3-phosphorylated Httex1. Httex1 with polyQ repeats below the pathogenic threshold (Httex1-23Q) was shown to aggregate in vitro and phosphorylation at T3 slows aggregation. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text