Chemical Synthesis of the Highly Hydrophobic Antiviral Membrane-Associated Protein IFITM3 and Modified Variants
Thibault J Harmand1, Vijaya R Pattabiraman1, Jeffrey W Bode1,2
1Laboratorium für Organische Chemie Department of Chemistry and Applied Biosciences ETH Zürich Wolfgang Pauli Strasse 10 8093 Zürich Switzerland.
Abstract:
Interferon-induced transmembrane protein 3 (IFITM3) is an antiviral transmembrane protein that is thought to serve as the primary factor for inhibiting the replication of a large number of viruses, including West Nile virus, Dengue virus, Ebola virus, and Zika virus. Production of this 14.5 kDa, 133-residue transmembrane protein, especially with essential posttranslational modifications, by recombinant expression is challenging. In this report, we document the chemical synthesis of IFTIM3 in multi-milligram quantities (>15 mg) and the preparation of phosphorylated and fluorescent variants. The synthesis was accomplished by using KAHA ligations, which operate under acidic aqueous/organic mixtures that excel at solubilizing even the exceptionally hydrophobic C-terminal region of IFITM3. The synthetic material is readily incorporated into model vesicles and forms the basis for using synthetic, homogenous IFITM3 and its derivatives for further studying its structure and biological mode of action.
Insights
Chemically synthesizing the antiviral protein Interferon-induced transmembrane protein 3 (IFITM3) in large quantities is now possible. This breakthrough enables further study of IFITM3
Area of Science:
- Biochemistry
- Molecular Biology
- Virology
Background:
- Interferon-induced transmembrane protein 3 (IFITM3) is a key antiviral protein.
- IFITM3 inhibits replication of viruses like Dengue, Ebola, and Zika.
- Recombinant production of IFITM3, especially with modifications, is difficult.
Purpose of the Study:
- To chemically synthesize multi-milligram quantities of IFITM3.
- To prepare phosphorylated and fluorescent variants of IFITM3.
- To enable further structural and mechanistic studies of IFITM3.
Main Methods:
- Chemical synthesis of IFITM3 using KAHA ligations.
- Utilizing acidic aqueous/organic mixtures for synthesis.
- Preparation of modified IFITM3 variants (phosphorylated, fluorescent).
Main Results:
- Successful synthesis of >15 mg of IFITM3.
- Developed a method to handle the hydrophobic C-terminal region.
- Created phosphorylated and fluorescent IFITM3 derivatives.
- Synthetic IFITM3 integrates into model vesicles.
Conclusions:
- Chemical synthesis provides a viable route for producing homogenous IFITM3.
- Synthetic IFITM3 and its derivatives are valuable tools for research.
- This work facilitates deeper understanding of IFITM3's antiviral mechanisms.
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