Customized peptidoglycan surfaces to investigate innate immune recognition via surface plasmon resonance.

Customized peptidoglycan surfaces to investigate innate immune recognition via surface plasmon resonance.
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DOI:
10.1016/bs.mie.2021.12.004
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发表时间:
2022
影响因子:
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通讯作者:
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中科院分区:
生物学4区
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糖蛋白相互作用促进了细胞内和细胞间的一些最重要的生物分子过程。它们参与了不同的细胞途径,细胞间的相互作用,并与许多疾病有关,使这些相互作用引起了极大的兴趣。然而,它们的结构和功能的多样性给在分子水平上的研究带来了巨大的挑战。表面等离子体共振(SPR)技术因其优越的灵敏度、实时监测相互作用的能力、相对简单的数据解释以及最重要的是无需荧光标记而直接测量结合而在研究糖-蛋白质相互作用方面显示出巨大的优势。在这里,SPR在研究糖蛋白相互作用中的另一个维度是通过人的先天免疫受体与它们的细菌肽多糖配体之间的结合的例子来证明的。为了更好地模拟溶液中的相互作用,采用了一种新的策略,将不同位置的碳水化合物拴在生物传感器表面,以表示碳水化合物配体对受体的潜在展示。随后的动力学分析提供了对与其受体结合的肽聚糖片段的优化配置的见解。该手稿包含一份“如何操作指南”,以帮助在其他糖蛋白结合系统中实施这些方法。
Glycan-protein interactions facilitate some of the most important biomolecular processes in and between cells. They are involved in different cellular pathways, cell-cell interactions and associated with many diseases, making these interactions of great interest. However, their structural and functional diversity poses great challenges in studying them at the molecular level. Surface plasmon resonance (SPR) technology presents great advantages to study glycan-protein interactions due to its superior sensitivity, ability to monitor real-time interactions, relatively simple data interpretation, and most importantly, direct measurement of binding without a need for fluorescent labeling. Here, another dimensionality of SPR in studying glycan-protein interactions is demonstrated via examples of binding between human innate immune receptors and their bacterial peptidoglycan ligands. In order to best resemble interactions in solution, a novel strategy of tethering the carbohydrate at different positions to the biosensor surface is applied to represent the potential displays of the carbohydrate ligand to the receptor. Subsequent kinetic analysis provides insights into the optimized configuration of peptidoglycan fragments for binding with its receptors. The manuscript contains a "how-to guide" to help with the implementation of these methods in other glycan-protein binding systems.
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发表时间: 2012-08-22
影响因子: 15
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影响因子: 64.5
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