CPMAS NMR platform for direct compositional analysis of mycobacterial cell-wall complexes and whole cells.
CPMAS NMR platform for direct compositional analysis of mycobacterial cell-wall complexes and whole cells.
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CPMAS NMR 平台，用于直接对分枝杆菌细胞壁复合物和全细胞进行成分分析。
DOI：
10.1016/j.jmro.2023.100127
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发表时间：
2023
影响因子：
--
通讯作者：
Cegelski,Lynette
中科院分区：
文献类型：
--
作者：
Liu,Xinyu;Brčić,Jasna;Cassell,GailH;Cegelski,Lynette
结核病和非结核分枝杆菌感染每年都在上升，往往导致慢性不治之症。重要的抗生素靶向细胞壁的生物合成，但一些分枝杆菌是惊人的耐药或耐受目前可用的抗生素。这种耐药性通常归因于不同分枝杆菌菌株和物种的复杂细胞壁组成的假定差异。然而，由于分枝杆菌细胞壁的高度交联和不溶性的性质，直接比较测定细胞壁的组成构成了挑战，通过传统的生化分析分析。我们介绍了一种方法来直接观察的化学成分的分枝杆菌细胞壁使用固态NMR光谱。13 C CPMAS光谱提供的单个组件（肽聚糖，阿拉伯半乳聚糖，和分枝菌酸）和ofin situcell-wall复合物。我们分配了细胞壁光谱中每个组分的光谱贡献。我们发现M细胞壁中阿拉伯半乳聚糖与肽聚糖的比例较高。一种以其抗生素抗性而闻名的微生物，相对于M。恶臭此外，不同类型的细胞壁靶向抗生素的差异化的影响，观察到在光谱的抗生素处理的全细胞。该平台将在评估不同分枝杆菌的细胞壁组成和抗生素活性以及考虑最有效的联合治疗方案方面具有价值。
Tuberculosis and non-tuberculosis mycobacterial infections are rising each year and often result in chronic incurable disease. Important antibiotics target cell-wall biosynthesis, yet some mycobacteria are alarmingly resistant or tolerant to currently available antibiotics. This resistance is often attributed to assumed differences in composition of the complex cell wall of different mycobacterial strains and species. However, due to the highly crosslinked and insoluble nature of mycobacterial cell walls, direct comparative determinations of cell-wall composition pose a challenge to analysis through conventional biochemical analyses. We introduce an approach to directly observe the chemical composition of mycobacterial cell walls using solid-state NMR spectroscopy.13C CPMAS spectra are provided of individual components (peptidoglycan, arabinogalactan, and mycolic acids) and ofin situcell-wall complexes. We assigned the spectroscopic contributions of each component in the cell-wall spectrum. We uncovered a higher arabinogalactan-to-peptidoglycan ratio in the cell wall ofM. abscessus, an organism noted for its antibiotic resistance, relative toM. smegmatis. Furthermore, differentiating influences of different types of cell-wall targeting antibiotics were observed in spectra of antibiotic-treated whole cells. This platform will be of value in evaluating cell-wall composition and antibiotic activity among different mycobacteria and in considering the most effective combination treatment regimens.