有机化学 ›› 2024, Vol. 44 ›› Issue (7): 2296-2304.DOI: 10.6023/cjoc202402006 上一篇    下一篇

研究论文

糖基化干扰素-γ的半合成

周敏园a, 赵洁b, 叶发荣b, 黄平b, 邓明刚a,*(), 王平b,*()   

  1. a 华中农业大学生命科学与技术学院 武汉 430070
    b 上海交通大学 张江高等研究院 上海手性药物分子工程重点实验室 化学糖生物学中心 上海 200240
  • 收稿日期:2024-02-05 修回日期:2024-03-17 发布日期:2024-04-10
  • 作者简介:
    † 共同第一作者.
  • 基金资助:
    国家自然科学基金(21672146); 国家自然科学基金(22077080); 国家自然科学基金(92253302); 上海交通大学医工交叉基金(21TQ1400210)

Semi-synthesis of Glycosylated Interferon-γ

Minyuan Zhoua, Jie Zhaob, Farong Yeb, Ping Huangb, Minggang Denga(), Ping Wangb()   

  1. a College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070
    b Center for Chemical Glycobiology, Shanghai Key Laboratory for Molecular Engineering of Chiral Drugs, Zhangjiang Institute for Advanced Study, Shanghai Jiao Tong University, Shanghai 200240
  • Received:2024-02-05 Revised:2024-03-17 Published:2024-04-10
  • Contact: E-mail: wangp1@sjtu.edu.cn;dengmg@mail.hzau.edu.cn
  • About author:
    † These authors contributed equally to this work.
  • Supported by:
    National Natural Science Fund(21672146); National Natural Science Fund(22077080); National Natural Science Fund(92253302); Cross Fund of Shanghai Jiao Tong University Medical Engineering(21TQ1400210)

干扰素-γ (IFN-γ)是一种重要的细胞因子, 具有增强免疫活性、抗肿瘤和抗病毒的作用, 在医学研究和临床应用中具有广泛的潜力. 本研究报道了一种均一N-GlcNAc修饰的IFN-γ的化学半合成方法. 利用化学方法合成制备糖肽片段(Pyr1-Leu33)和多肽片段(Lys34-Ser39), 并通过大肠杆菌表达得到多肽片段(Ser40-Gly138). 采用“表达丝氨酸连接”和“自然化学连接-脱硫”相结合的策略, 成功完成了自C端到N端的N-GlcNAc修饰的IFN-γ蛋白的合成, 并成功地对目标糖蛋白进行了复性.

关键词: 自然化学连接, 表达丝氨酸连接, β-硫代氨基酸, 糖基化修饰

Interferon-gamma (IFN-γ) is an important cytokine with enhanced immune activity, anti-tumor and antiviral effects, and holds significant potential in medical research and clinical applications. In this study, we report a highly efficient semi-synthesis strategy of homogeneous N-GlcNAc modified IFN-γ. The glycopeptide fragment (Pyr1-Leu33) and peptide fragment (Lys34-Ser39) were prepared through chemical methods. And the peptide fragment (Ser40-Gly138) was obtained through Escherichia coli (E. coli) expression. Subsequently, using a combination of “expressed serine ligation” and “native chemical ligation-desulfurization”, we ligated these fragments from the C-terminal to the N-terminal, resulting in a full-length glycoprotein, which was successfully refolded to obtain the desired product.

Key words: native chemical ligation, expressed serine ligation, β-thioamino acid, glycosylation