Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Am Mühlenberg 1, 14476, Potsdam, Germany.
Department of Chemistry and Biochemistry, Freie Universität Berlin, Arnimallee 22, 14195, Berlin, Germany.
Angew Chem Int Ed Engl. 2021 Jun 7;60(24):13302-13309. doi: 10.1002/anie.202102690. Epub 2021 May 7.
Protein-glycan interactions mediate important biological processes, including pathogen host invasion and cellular communication. Herein, we showcase an expedite approach that integrates automated glycan assembly (AGA) of F-labeled probes and high-throughput NMR methods, enabling the study of protein-glycan interactions. Synthetic Lewis type 2 antigens were screened against seven glycan binding proteins (GBPs), including DC-SIGN and BambL, respectively involved in HIV-1 and lung infections in immunocompromised patients, confirming the preference for fucosylated glycans (Le , H type 2, Le ). Previously unknown glycan-lectin weak interactions were detected, and thermodynamic data were obtained. Enzymatic reactions were monitored in real-time, delivering kinetic parameters. These results demonstrate the utility of AGA combined with F NMR for the discovery and characterization of glycan-protein interactions, opening up new perspectives for F-labeled complex glycans.
蛋白-聚糖相互作用介导着重要的生物学过程，包括病原体宿主入侵和细胞通讯。在此，我们展示了一种集成的方法，该方法整合了 F 标记探针的自动化聚糖组装（AGA）和高通量 NMR 方法，从而能够研究蛋白-聚糖相互作用。合成的 Lewis 型 2 抗原被筛选出与 7 种糖结合蛋白（GBP）相互作用，包括 DC-SIGN 和 BambL，分别涉及 HIV-1 和免疫功能低下患者肺部感染，证实了对岩藻糖基化聚糖（Le, H 型 2, Le ）的偏好。检测到先前未知的聚糖-凝集素弱相互作用，并获得热力学数据。实时监测酶反应，提供动力学参数。这些结果证明了 AGA 与 19 F NMR 结合用于发现和表征糖蛋白相互作用的效用，为 19 F 标记的复杂聚糖开辟了新的视角。
