Single-Cell Force Spectroscopy of Als-Mediated Fungal Adhesion.
Single-Cell Force Spectroscopy of Als-Mediated Fungal Adhesion.
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DOI：
10.1039/c3ay40473k
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发表时间：
2013-08-07
期刊：
影响因子：
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通讯作者：
Dufrêne YF
中科院分区：
文献类型：
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作者：
Alsteens D;Beaussart A;Derclaye S;El-Kirat-Chatel S;Park HR;Lipke PN;Dufrêne YF
传统上用于研究微生物细胞粘附行为的宏观测定提供了在大量细胞上获得的平均信息，并且不测量驱动单细胞粘附的基本力。在这里，我们使用单细胞力谱（SCFS）来量化参与人类真菌病原体白色念珠菌粘附的特异性和非特异性力。使用仿生聚多巴胺湿聚合物将表达白色念珠菌粘附蛋白 Als5p 的酿酒酵母细胞附着在原子力显微镜无尖悬臂上，并记录获得的细胞探针和各种固体表面之间的力-距离曲线。在疏水基底上获得的力特征表现出较大的粘附力（1.25±0.2 nN）和延长的断裂长度（高达 400 nm），这归因于 Als5p 的疏水串联重复的结合和拉伸。在纤连蛋白 (Fn) 涂覆的基质上收集的数据具有很强的粘附力 (2.8 ± 0.6 nN)，反映了 Fn 与 Als5p 的 N 端免疫球蛋白样区域之间的特异性结合，随后是弱粘附性大分子键。疏水性和 Fn 粘附力都随着接触时间的增加而增加，强调了时间在增强粘附力方面的重要作用。我们的 SCFS 方法为生物医学提供了一个多功能平台，用于了解驱动真菌病原体粘附和生物膜形成的基本力量。
Macroscopic assays that are traditionally used to investigate the adhesion behaviour of microbial cells provide averaged information obtained on large populations of cells and do not measure the fundamental forces driving single-cell adhesion. Here, we use single-cell force spectroscopy (SCFS) to quantify the specific and non-specific forces engaged in the adhesion of the human fungal pathogen Candida albicans. Saccharomyces cerevisiae cells expressing the C. albicans adhesion protein Als5p were attached on atomic force microscopy tipless cantilevers using a bioinspired polydopamine wet polymer, and force-distance curves were recorded between the obtained cell probes and various solid surfaces. Force signatures obtained on hydrophobic substrates exhibited large adhesion forces (1.25 ± 0.2 nN) with extended rupture lengths (up to 400 nm), attributed to the binding and stretching of the hydrophobic tandem repeats of Als5p. Data collected on fibronectin (Fn) -coated substrates featured strong adhesion forces (2.8 ± 0.6 nN), reflecting specific binding between Fn and the N-terminal immunoglobulin-like regions of Als5p, followed by weakly adhesive macromolecular bonds. Both hydrophobic and Fn adhesion forces increased with contact time, emphasizing the important role that time plays in strengthening adhesion. Our SCFS methodology provides a versatile platform in biomedicine for understanding the fundamental forces driving adhesion and biofilm formation in fungal pathogens.