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实现蛋白质-聚糖相互作用的高通量NMR研究
作者:小柯机器人 发布时间:2021/4/4 23:15:32

德国马克斯·普朗克研究所Martina Delbianco团队实现了19F标记的聚糖探针的自动聚糖组装,使蛋白质聚糖相互作用的高通量NMR研究成为可能。相关研究成果于2021年3月31日发表于国际一流学术期刊《德国应用化学》。

研究人员展示了一种快速的方法,该方法集成了19个F标记探针的自动聚糖组装(AGA)和高通量NMR方法,使蛋白质-聚糖相互作用的研究成为可能。合成的Lewis 2型抗原针对7种聚糖结合蛋白(GBPs)进行筛选,包括DC-SIGN和BambL,分别参与免疫功能低下患者的HIV-1和肺部感染,证实了对岩藻糖基聚糖(Lex,H 2型,Ley)的偏好。

研究检测到以前未知的聚糖-凝集素弱相互作用,并获得热力学数据。实时监测酶促反应获得了动力学参数。

研究结果证明了AGA结合19 F NMR在发现和表征聚糖-蛋白质相互作用方面的实用性,为19 F标记的复杂聚糖开辟了新的前景。

附:英文原文

Title: Automated glycan assembly of 19F labelled glycan probes enables high‐throughput NMR studies of protein‐glycan interactions

Author: Giulio Fittolani, Elena Shanina, M, Peter H. Seeberger, Christoph Rademacher, Martina Delbianco

Issue&Volume: 2021-03-31

Abstract: Protein‐glycan interactions mediate important biological processes, including pathogen host invasion and cellular communication. Major challenges to monitoring these low affinity interactions are the required high sensitivity of a biophysical assay and to cover a breath of synthetic well‐defined structures. Here, we showcase an expedite approach that integrates automated glycan assembly (AGA) of  19  F labelled 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  x  , H type 2, Le  y  ). 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  19  F NMR for the discovery and characterization of glycan‐protein interactions, opening up new perspectives for  19  F labelled complex glycans.

DOI: 10.1002/anie.202102690

Source: https://onlinelibrary.wiley.com/doi/10.1002/anie.202102690

期刊信息

Angewandte Chemie:《德国应用化学》,创刊于1887年。隶属于德国化学会,最新IF:12.959
官方网址:https://onlinelibrary.wiley.com/journal/15213773
投稿链接:https://www.editorialmanager.com/anie/default.aspx