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.
Das hydrophobe antivirale Membranprotein IFITM3 wurde durch chemoselektive Ligation unter sauren Bedingungen aufgebaut; dabei werden Estergruppen eingeführt, welche die Löslichkeit der Peptidsegmente erhöhen. Durch α‐Ketosäure‐Hydroxylamin(KAHA)‐Ligation erhaltene Depsipeptide wurden am Ende der Proteinsynthese in Amide umgelagert.
Keywords: Chemische Proteinsynthese; IFITM3; KAHA-Ligation; Membranproteine; Proteinmodifikationen.
© 2017 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.