Francius Grégory, Petit Florian, Clément Eloïse, Chekli Yankel, Ghigo Jean-Marc, Beloin Christophe, Duval Jérôme F L
Université de Lorraine, LCPME, UMR 7564, Villers-lès-Nancy, F-54600, France.
Nanoscale. 2021 Jan 14;13(2):1257-1272. doi: 10.1039/d0nr06840c. Epub 2021 Jan 6.
Yad fimbriae are currently viewed as versatile bacterial adhesins able to significantly mediate host or plant-pathogen recognition and contribute to the persistence of Escherichia coli in both the environment and within hosts. To date, however, the underlying adhesion process of Yad fimbriae on surfaces defined by controlled coating chemistries has not been evaluated on the relevant molecular scale. In this work, the interaction forces operational between Yad fimbriae expressed by genetically modified E. coli and self-assembled monolayers (SAM) differing in terms of charge, hydrophobicity or the nature of decorating sugar units are quantified by Single Molecule Force Spectroscopy (SMFS) on the nanoscale. It is found that the adhesion of Yad fimbriae onto probes functionalized with xylose is as strong as that measured with probes decorated with anti-Yad antibodies (ca. 80 to 300 pN). In contrast, the interactions of Yad with galactose, lactose, mannose, -OH, -NH, -COOH and -CH terminated SAMs are clearly non-specific. Interpretation of SMFS measurements on the basis of worm-like-chain modeling for polypeptide nanomechanics further leads to the estimates of the maximal extension of Yad fimbriae upon stretching, of their persistence length and of their polydispersity. Finally, we show for the first time a strong correlation between the adhesion properties of Yad-decorated bacteria determined from conventional macroscopic counting methods and the molecular adhesion capacity of Yad fimbriae. This demonstration advocates the effort that should be made to understand on the nanoscale level the interactions between fimbriae and their cognate ligands. The results could further help the design of potential anti-adhesive molecules or surfaces to better fight against the virulence of bacterial pathogens.
Yad菌毛目前被视为多功能细菌粘附素,能够显著介导宿主或植物与病原体的识别,并有助于大肠杆菌在环境和宿主体内的持续存在。然而,迄今为止,尚未在相关分子尺度上评估Yad菌毛在由可控涂层化学定义的表面上的潜在粘附过程。在这项工作中,通过单分子力谱(SMFS)在纳米尺度上对经基因改造的大肠杆菌表达的Yad菌毛与在电荷、疏水性或修饰糖单元性质方面不同的自组装单分子层(SAM)之间的相互作用力进行了量化。研究发现,Yad菌毛与用木糖功能化的探针的粘附力与用抗Yad抗体修饰的探针所测得的粘附力一样强(约80至300 pN)。相比之下,Yad与半乳糖、乳糖、甘露糖、-OH、-NH、-COOH和-CH终止的SAM之间的相互作用显然是非特异性的。基于多肽纳米力学的蠕虫状链模型对SMFS测量结果的解释进一步得出了Yad菌毛在拉伸时的最大伸展长度、其持续长度及其多分散性的估计值。最后,我们首次表明,从传统宏观计数方法确定的Yad修饰细菌的粘附特性与Yad菌毛的分子粘附能力之间存在很强的相关性。这一证明提倡应努力在纳米尺度上理解菌毛与其同源配体之间的相互作用。这些结果可能进一步有助于设计潜在的抗粘附分子或表面,以更好地对抗细菌病原体的毒力。