Systematic Assessment of Accessibility to the Surface of Staphylococcus aureus.

Systematic Assessment of Accessibility to the Surface of Staphylococcus aureus.
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DOI:
10.1021/acschembio.1c00604
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发表时间:
2021-11-19
影响因子:
4
通讯作者:
Pires, Marcos M.
Pires, Marcos M.
中科院分区:
生物学2区
文献类型:
--
作者:
Ferraro, Noel J.;Kim, Seonghoon;Im, Wonpil;Pires, Marcos M.

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来自细菌敌人、抗微生物肽和宿主免疫蛋白的蛋白质必须穿过细菌表面生物大分子的致密层到达革兰氏阳性细菌的肽聚糖(PG)层。分子的亚类(例如,具有细胞内靶标的抗生素)也必须渗透通过PG(以分子筛方式)以到达细胞质膜。尽管表面可及性的生物学和治疗的重要性,在活细菌细胞的系统分析一直缺乏。我们描述了一种活细胞荧光测定法,该测定法是稳健的,显示出高水平的再现性,并报告了分子对PG支架的渗透性和PG支架内的渗透性。此外,我们的研究表明,磷壁酸阻碍了各种大小和化学成分的分子的渗透性。细菌细胞表面可及性是许多抗生素和免疫蛋白的关键。我们描述了一种系统地报告分子渗透到细菌细胞壁的测定。
Proteins from bacterial foes, antimicrobial peptides, and host immune proteins must navigate past a dense layer of bacterial surface biomacromolecules to reach the peptidoglycan (PG) layer of Gram-positive bacteria. A subclass of molecules (e.g., antibiotics with intracellular targets) also must permeate through the PG (in a molecular sieving manner) to reach the cytoplasmic membrane. Despite the biological and therapeutic importance of surface accessibility, systematic analyses in live bacterial cells have been lacking. We describe a live cell fluorescence assay that is robust, shows high level of reproducibility, and reports on the permeability of molecules to and within the PG scaffold. Moreover, our study shows that teichoic acids impede the permeability of molecules of a wide range of sizes and chemical composition. Bacterial cell surface accessibility is key for many antibiotics and immune proteins. We describe an assay that systematically reports on permeation of molecules to the cell wall of bacteria.
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