用于分枝杆菌细胞壁复合物和整个细胞直接成分分析的CPMAS核磁共振平台

IF 2.624
Xinyu Liu , Jasna Brčić , Gail H. Cassell , Lynette Cegelski
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引用次数: 1

摘要

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生物合成,然而一些分枝杆菌对目前可用的抗生素具有惊人的耐药性或耐受性。这种耐药性通常归因于不同分枝杆菌菌株和种类的复杂细胞壁组成的假设差异。然而,由于分枝杆菌细胞壁的高度交联和不溶性,细胞壁组成的直接比较测定对传统的生化分析提出了挑战。我们介绍了一种利用固态核磁共振光谱直接观察分枝杆菌细胞壁化学成分的方法。13C CPMAS光谱提供了单个成分(肽聚糖,阿拉伯半乳糖和霉菌酸)和原位细胞壁复合物。我们分配了细胞壁光谱中每个组分的光谱贡献。我们发现在脓疡分枝杆菌的细胞壁中,阿拉伯半乳糖与肽聚糖的比例更高,这是一种以抗生素耐药性而闻名的有机体,相对于耻垢分枝杆菌。此外,在抗生素处理的全细胞光谱中观察到不同类型的细胞壁靶向抗生素的差异影响。该平台将在评估不同分枝杆菌之间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。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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

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 of in situ cell-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 of M. abscessus, an organism noted for its antibiotic resistance, relative to M. 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.

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1.90
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