Mass Spectrometry of Biomacromolecules and ‡Molecular Biology & Microbial Food Safety, Swammerdam Institute for Life Sciences, University of Amsterdam , Amsterdam, The Netherlands.
J Proteome Res. 2013 Oct 4;12(10):4507-21. doi: 10.1021/pr4005629. Epub 2013 Sep 23.
Bacillus cereus, responsible for food poisoning, and Clostridium difficile, the causative agent of Clostridium difficile-associated diarrhea (CDAD), are both spore-forming pathogens involved in food spoilage, food intoxication, and other infections in humans and animals. The proteinaceous coat and the exosporium layers from spores are important for their resistance and pathogenicity characteristics. The exosporium additionally provides an ability to adhere to surfaces eventually leading to spore survival in food. Thus, studying these layers and identifying suitable protein targets for rapid detection and removal of spores is of the utmost importance. In this study, we identified 100 proteins from B. cereus spore coat, exosporium and 54 proteins from the C. difficile coat insoluble protein fraction. In an attempt to define a universal set of spore outer layer proteins, we identified 11 superfamily domains common to the identified proteins from two Bacilli and one Clostridium species. The evaluated orthologue relationships of identified proteins across different spore formers resulted in a set of 13 coat proteins conserved across the spore formers and 12 exosporium proteins conserved in the B. cereus group, which could be tested for quick and easy detection or targeted in strategies aimed at removal of spores from surfaces.
蜡样芽胞杆菌可引起食物中毒，艰难梭菌是导致艰难梭菌相关性腹泻（CDAD）的病原体，两者均为形成孢子的病原体，可引起人类和动物的食物腐败、食物中毒和其他感染。孢子的蛋白质外壳和外孢囊层对于其耐药性和致病性特征很重要。外孢囊层还提供了一种粘附表面的能力，最终导致孢子在食物中存活。因此，研究这些层并确定适合快速检测和去除孢子的合适蛋白质靶标非常重要。在这项研究中，我们从蜡样芽胞杆菌孢子外壳、外孢囊层中鉴定了 100 种蛋白质，从艰难梭菌外壳不溶性蛋白部分中鉴定了 54 种蛋白质。为了定义一套通用的孢子外层蛋白，我们鉴定了来自两种芽孢杆菌和一种梭菌的鉴定蛋白中 11 个超家族结构域。在不同孢子形成体之间鉴定蛋白质的直系同源物关系，产生了一套在孢子形成体中保守的 13 种外壳蛋白和在芽孢杆菌组中保守的 12 种外孢囊蛋白，这些蛋白可以用于快速、简便的检测，或者作为从表面去除孢子的靶向策略的目标。
