北京大学董甦伟研究团队报道了N -聚糖在糖蛋白化学合成高级糖基化终产物V结构域受体中的功能研究。相关研究成果发表在2024年6月25日出版的《美国化学会杂志》。

晚期糖基化终产物受体（RAGE）在炎症相关途径和各种慢性疾病中发挥着至关重要的作用。尽管在RAGE的配体结合V结构域（VD）中N-糖基化具有公认的重要性，但由于通过生物表达获得同质糖蛋白样品的挑战，缺乏对位点活性和结构-活性关系的全面了解。

该文中，研究人员结合化学和化学酶法，通过结合精确的N-聚糖结构，合成了具有Asn3糖基化的RAGE-VD及其同源物。对这些样品的评估表明，与其他RAGE-VD形式相比，Asn3位点的α2,6-唾液酸化N-糖基化对RAGE过表达细胞中HMGB1诱导的核因子-κB（NF-κB）表达产生更有效的抑制作用。

氢/氘交换质谱分析显示HMGB1内存在唾液酸化RAGE VD诱导的相互作用区。相反，VD中的Asn3-N-糖基化对RAGE-VD/S100B相互作用的影响可以忽略不计。

该研究建立了一种获得均匀糖基化的RAGE-VD的方法，并探讨了N-糖基化对RAGE-VD及其配体蛋白之间相互作用的调节作用。

附：英文原文

Title: Investigation of N-Glycan Functions in Receptor for Advanced Glycation End Products V Domain through Chemical Glycoprotein Synthesis

Author: Weidong Dong, Xingyue Yang, Xinyu Li, Sheng Wei, Chuanjing An, Jun Zhang, Xiaomeng Shi, Suwei Dong

Issue&Volume: June 25, 2024

Abstract: The receptor for advanced glycation end products (RAGE) plays a crucial role in inflammation-related pathways and various chronic diseases. Despite the recognized significance of N-glycosylation in the ligand-binding V domain (VD) of RAGE, a comprehensive understanding of the site-activity and structure–activity relationships is lacking due to the challenges in obtaining homogeneous glycoprotein samples through biological expression. Here, we combined chemical and chemoenzymatic approaches to synthesize RAGE-VD and its congeners with Asn3-glycosylation by incorporating precise N-glycan structures. Evaluation of these samples revealed that, in comparison to other RAGE-VD forms, α2,6-sialylated N-glycosylation at the Asn3 site results in more potent inhibition of HMGB1-induced nuclear factor-κB (NF-κB) expression in RAGE-overexpressing cells. Hydrogen/deuterium exchange-mass spectrum analysis revealed a sialylated RAGE-VD-induced interaction region within HMGB1. Conversely, Asn3 N-glycosylation in VD has negligible effects on RAGE-VD/S100B interactions. This study established an approach for accessing homogeneously glycosylated RAGE-VD and explored the modulatory effects of N-glycosylation on the interactions between RAGE-VD and its ligand proteins.

DOI: 10.1021/jacs.4c01413

Source: https://pubs.acs.org/doi/abs/10.1021/jacs.4c01413