均一糖基化修饰白细胞介素-10的化学合成研究  

Studies toward Chemical Synthesis of Homogeneously Glycosylated Interleukin-10

作　　者：董伟东 刘海韵 李信宇 王标 成波 董甦伟 Weidong Dong;Haiyun Liu;Xinyu Li;Biao Wang;Bo Cheng;Suwei Dong(State Key Laboratory of Natural and Biomimetic Drugs,Chemical Biology Center,and Department of Chemical Biology at School of Pharmaceutical Sciences,Peking University,Beijing 100191)

机构地区：[1]北京大学药学院化学生物学系、化学生物学中心、天然药物及仿生药物全国重点实验室,北京100191

出　　处：《有机化学》2025年第3期951-958,共8页Chinese Journal of Organic Chemistry

基　　金：基金资助:国家自然科学基金(92153301);国家自然科学基金(22321005)。

摘　　要：细胞因子人白细胞介素-10 (IL-10)序列中含有一个保守的天冬酰胺糖基化(N-糖基化)基序, 但尚无相关研究报道这一糖基化的存在与潜在功能. 探索了IL-10及其糖基化形式的化学合成与表征: 基于固相多肽合成、自然化学连接和无金属巯基脱除等策略, 实现IL-10全长序列的构建, 并通过优化折叠复性条件成功获得未经糖基修饰的IL-10. 在此合成路线基础上, 利用糖修饰氨基酸砌块插入策略获得N-乙酰葡糖胺(N-GlcNAc)修饰的糖多肽片断, 进一步获得了天冬酰胺116位点单糖修饰的IL-10. 最后, 经过对合成IL-10样品的质谱和圆二色谱评价, 证明其具有与生物表达样品相似的结构.Human cytokine interleukin-10 (IL-10) sequence contains a conserved N-glycosylation sequon, Asn-Lys-Ser, but the presence and functional significance of this glycosylation have not been previously studied. Here, the chemical synthesis and characterization of IL-10 and its glycosylated form were explored. Using solid-phase peptide synthesis, native chemical ligation, and metal-free desulfurization, the full-length IL-10 was assembled and the non-glycosylated protein was successfully obtained through optimizing the refolding conditions. Building on this synthetic route, an N-acetylglucosamine (N-GlcNAc) modification was introduced into the peptide segment via cassette strategy, and further elaborated into Asn116 N-mono- saccharide-modified IL-10. Mass spectrometry (MS) and circular dichroism analysis of the synthesized IL-10 variants have suggested that they have similar structures to that of the biologically expressed counterpart.

关 键 词：蛋白质化学合成 糖蛋白 自然化学连接 蛋白质折叠 人白细胞介素-10 

分 类 号：Q51[生物学—生物化学]