文献类型： 外文期刊

第一作者： Delcourte, Loic

作者： Delcourte, Loic;Berbon, Melanie;Lends, Alons;Grelard, Axelle;Habenstein, Birgit;Loquet, Antoine;Rodriguez, Marion;Delhaes, Laurence;Subban, Kamalraj;Daskalov, Asen;Morvan, Estelle;Saupe, Sven J.;Delhaes, Laurence;Aimanianda, Vishukumar;Daskalov, Asen;Daskalov, Asen;Loquet, Antoine

作者机构：

期刊名称：METHODS （ 影响因子：4.2； 五年影响因子：3.8 ）

ISSN： 1046-2023

年卷期： 2024 年 230 卷

页码：

收录情况： SCI

摘要： Most bacterial, plant and fungal cells possess at their surface a protective layer called the cell wall, conferring strength, plasticity and rigidity to withstand the osmotic pressure. This molecular barrier is crucial for pathogenic microorganisms, as it protects the cell from the local environment and often constitutes the first structural component encountered in the host-pathogen interaction. In pathogenic molds and yeasts, the cell wall constitutes the main target for the development of clinically-relevant antifungal drugs. In the past decade, solid-state NMR has emerged as a powerful analytical technique to investigate the molecular organization of microbial cell walls in the context of intact cells. 13 C NMR chemical shift is an exquisite source of information to identify the polysaccharides present in the cell wall, and two-dimensional 13 C- 13 C correlation experiments provide an efficient tool to rapidly access the polysaccharide composition in whole cells. Here we investigate the use of the adiabatic DREAM (for dipolar recoupling enhancement through amplitude modulation) recoupling scheme to improve solid-state NMR analysis of polysaccharides in intact cells. We demonstrate the advantages of two-dimensional 13 C- 13 C experiments using the DREAM recoupling scheme. We report the spectral editing of polysaccharide signals by varying the radio-frequency carrier position. We provide practical considerations for the implementation of DREAM experiments to characterize polysaccharides in whole cells. We demonstrate the approach on intact fungal cells of Neurospora crassa and Aspergillus fumigatus, , a model and a pathogenic filamentous fungus, respectively. The approach could be envisioned to efficiently reduce the spectral crowding of more complex cell surfaces, such as cell wall and peptidoglycan in bacteria.

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