Chemical Synthesis of the Highly Hydrophobic Antiviral Membrane Associated Protein IFITM3 and Modified Variants
 

Chemical Synthesis of the Highly Hydrophobic Antiviral Membrane Associated Protein IFITM3 and Modified Variants


Interferon-induced transmembrane protein 3 (IFITM3) is an antiviral transmembrane protein thought to serve as the primary factor for inhibiting replication of a large number of viruses, including the West Nile virus, the Dengue virus, the Ebola virus, and the 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 multimilligram quantities (>15 mg) and the preparation of phosphorylated and fluorescent variants. The synthesis was accomplished by 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 readily incorporates in model vesicles and forms the basis for using synthetic, homogenous IFITM3 and its derivatives for further studying its structure and biological mode of action.

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