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活细胞上蛋白质特异性聚糖氧化的表面增强拉曼光谱成像,用于定量可视化病原体与细胞的相互作用。

SERS imaging of protein-specific glycan oxidation on living cells to quantitatively visualize pathogen-cell interactions.

作者信息

Wang Yuru, Wu Shan, Yang Yuanjiao, Yang Yuhui, Liu Huipu, Chen Yunlong, Ju Huangxian

机构信息

State Key Laboratory of Analytical Chemistry for Life Science, School of Chemistry and Chemical Engineering, Nanjing University Nanjing 210023 China

出版信息

Chem Sci. 2024 Feb 2;15(11):3901-3906. doi: 10.1039/d4sc00157e. eCollection 2024 Mar 13.

Abstract

Glycan oxidation on the cell surface occurs in many specific life processes including pathogen-cell interactions. This work develops a surface-enhanced Raman scattering (SERS) imaging strategy for quantitative monitoring of protein-specific glycan oxidation mediated pathogen-cell interactions by utilizing Raman reporter DTNB and aptamer co-assembled platinum shelled gold nanoparticles (Au@Pt-DTNB/Apt). Using () and MCF-7 cells as models, Au@Pt-DTNB/Apt can specifically bind to MUC1 protein on the cell surface containing heavy galactose (Gal) and -acetylgalactosamine (GalNAc) modification. When interacts with cells, the secreted galactose oxidase (GO) can oxidize Gal/GalNAc, and the generated reactive oxygen species (ROS) further oxidizes DTNB to produce TNB for greatly enhancing the SERS signal. This strategy can quantitatively visualize for the first time both the protein-specific glycan oxidation and the mediated pathogen-cell interactions, thus providing key quantitative information to distinguish and explore the pathogen-resistance and pharmacological mechanisms of different drugs.

摘要

细胞表面的聚糖氧化发生在许多特定的生命过程中,包括病原体与细胞的相互作用。这项工作开发了一种表面增强拉曼散射(SERS)成像策略,通过利用拉曼报告分子5,5'-二硫代双(2-硝基苯甲酸)(DTNB)和适配体共组装的铂壳金纳米颗粒(Au@Pt-DTNB/Apt),对蛋白质特异性聚糖氧化介导的病原体与细胞的相互作用进行定量监测。以()和MCF-7细胞为模型,Au@Pt-DTNB/Apt可以特异性结合到细胞表面含有重半乳糖(Gal)和N-乙酰半乳糖胺(GalNAc)修饰的粘蛋白1(MUC1)蛋白上。当()与细胞相互作用时,分泌的半乳糖氧化酶(GO)可以氧化Gal/GalNAc,产生的活性氧(ROS)进一步氧化DTNB生成TNB,从而极大地增强SERS信号。该策略首次能够定量可视化蛋白质特异性聚糖氧化以及介导的病原体与细胞的相互作用,从而提供关键的定量信息,以区分和探索不同药物的抗病原体和药理机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/32ac/10935716/4a0964da3c29/d4sc00157e-f1.jpg

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