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Analysis of Staphylococcus aureus wall teichoic acid glycoepitopes by Fourier Transform Infrared Spectroscopy provides novel insights into the staphylococcal glycocode

机译:傅里叶变换红外光谱法对金黄色葡萄球菌壁骨壁酸糖表位的分析提供了对葡萄球菌糖码的新见解

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摘要

Surface carbohydrate moieties are essential for bacterial communication, phage-bacteria and host-pathogen interaction. Most Staphylococcus aureus produce polyribitolphosphate type Wall teichoic acids (WTAs) substituted with α- and/or β-O-linked N-acetyl-glucosamine (α-/β-O-GlcNAc) residues. GlcNAc modifications have attracted particular interest, as they were shown to govern staphylococcal adhesion to host cells, to promote phage susceptibility conferring beta-lactam resistance and are an important target for antimicrobial agents and vaccines. However, there is a lack of rapid, reliable, and convenient methods to detect and quantify these sugar residues. Whole cell Fourier transform infrared (FTIR) spectroscopy could meet these demands and was employed to analyse WTAs and WTA glycosylation in S. aureus. Using S. aureus mutants, we found that a complete loss of WTA expression resulted in strong FTIR spectral perturbations mainly related to carbohydrates and phosphorus-containing molecules. We could demonstrate that α- or β-O-GlcNAc WTA substituents can be clearly differentiated by chemometrically assisted FTIR spectroscopy. Our results suggest that whole cell FTIR spectroscopy represents a powerful and reliable method for large scale analysis of WTA glycosylation, thus opening up a complete new range of options for deciphering the staphylococcal pathogenesis related glycocode.
机译:表面碳水化合物部分对于细菌交流,噬菌体-细菌和宿主-病原体相互作用至关重要。大多数金黄色葡萄球菌会产生被α-和/或β-O连接的N-乙酰基葡萄糖胺(α-/β-O-GlcNAc)残基取代的聚核糖核酸型墙体壁酸(WTA)。 GlcNAc修饰引起了人们的特别兴趣,因为它们被证明可以控制葡萄球菌与宿主细胞的粘附,促进赋予β-内酰胺抗性的噬菌体敏感性,并且是抗菌剂和疫苗的重要目标。但是,缺少快速,可靠和方便的方法来检测和定量这些糖残基。全细胞傅里叶变换红外光谱(FTIR)可以满足这些要求,并被用于分析金黄色葡萄球菌中的WTA和WTA糖基化。使用金黄色葡萄球菌突变体,我们发现WTA表达的完全丧失导致强烈的FTIR光谱扰动,主要与碳水化合物和含磷分子有关。我们可以证明,通过化学计量辅助FTIR光谱可以清楚地区分α-或β-O-GlcNAcWTA取代基。我们的结果表明,全细胞FTIR光谱法代表了一种强大且可靠的方法,可用于WTA糖基化的大规模分析,从而为解密与葡萄球菌致病相关的糖典开辟了一个全新的选择范围。

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