Laboratorium für Organische Chemie, Department of Chemistry and Applied Biosciences, ETH Zürich, Wolfgang Pauli Strasse 10, 8093, Zürich, Switzerland.
Institue of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Chisuka, Nagoya, 464-8602, Japan.
Angew Chem Int Ed Engl. 2017 Oct 2;56(41):12639-12643. doi: 10.1002/anie.201707554. Epub 2017 Sep 7.
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.
干扰素诱导跨膜蛋白 3（IFITM3）是一种抗病毒跨膜蛋白，被认为是抑制多种病毒复制的主要因素，包括西尼罗河病毒、登革热病毒、埃博拉病毒和寨卡病毒。通过重组表达生产这种 14.5 kDa、133 个氨基酸残基的跨膜蛋白，特别是具有必要的翻译后修饰，具有挑战性。在本报告中，我们记录了多毫克（>15 mg）数量的 IFITM3 的化学合成以及磷酸化和荧光变体的制备。该合成是通过使用 KAHA 连接来完成的，该连接在酸性水/有机溶剂混合物中进行，该混合物非常擅长溶解 IFITM3 异常疏水的 C 末端区域。合成材料很容易掺入模型囊泡中，并为使用合成的、均一的 IFITM3 及其衍生物进一步研究其结构和生物学作用模式奠定了基础。
