Sanchez Javier, Romero-Rodriguez Alba, Troncoso-Cotal Scarlett, Paredes-Sabja Daniel
bioRxiv. 2024 Dec 20:2024.04.07.588478. doi: 10.1101/2024.04.07.588478.
spores are essential for initiation, recurrence and transmission of the disease. The spore surface layers are composed of an outermost exosporium layer that surrounds another proteinaceous layer, the spore coat. These spore surfaces layers are responsible for initial interactions with the host and spore resistance properties contributing to transmission and recurrence of CDI. During spore-development, assembly of both layers is tightly interconnected thus studying the surface is essential for understanding the assembly of these layers and identification of potential targets for therapeutics. Several spore coat /exosporium extraction methods have utilized different extraction procedures making comparison across studies difficult and their impact on spore surface layer properties remains unclear. Here, we tested how commonly used chemical methods remove the spore coat /exosporium layers, analyzing treated-spores by phase contrast microscopy, transmission electron microscopy, western blotting, and lysozyme-triggered germination to functionally characterize the extraction efficiency of these treatment on these layers. Our results provide a systematic analysis and offer a platform for future spore coat and exosporium-related studies.
芽孢对于疾病的起始、复发和传播至关重要。芽孢表面层由最外层的芽孢外壁层组成，该外壁层围绕着另一个蛋白质层，即芽孢衣。这些芽孢表面层负责与宿主的初始相互作用以及有助于艰难梭菌感染传播和复发的芽孢抗性特性。在芽孢发育过程中，这两层的组装紧密相连，因此研究表面对于理解这些层的组装以及确定治疗的潜在靶点至关重要。几种芽孢衣/芽孢外壁提取方法采用了不同的提取程序，使得跨研究比较变得困难，并且它们对芽孢表面层特性的影响仍不清楚。在这里，我们测试了常用化学方法如何去除芽孢衣/芽孢外壁层，通过相差显微镜、透射电子显微镜、蛋白质免疫印迹和溶菌酶触发的萌发来分析处理后的芽孢，以从功能上表征这些处理对这些层的提取效率。我们的结果提供了系统分析，并为未来与芽孢衣和芽孢外壁相关的研究提供了一个平台。
