Surgical Infection Research Group, Faculty of Medicine, Health and Human Sciences, Macquarie University, Sydney 2109, Australia.
Australian Proteome Analysis Facility, Macquarie University, Sydney 2109, Australia.
Int J Mol Sci. 2022 Jun 8;23(12):6415. doi: 10.3390/ijms23126415.
is a notorious biofilm-producing pathogen that is frequently isolated from implantable medical device infections. As biofilm ages, it becomes more tolerant to antimicrobial treatment leading to treatment failure and necessitating the costly removal of infected devices. In this study, we performed in-solution digestion followed by TMT-based high-throughput mass spectrometry and investigated what changes occur in the proteome of biofilm grown for 3-days and 12-days in comparison with 24 h planktonic. It showed that proteins associated with biosynthetic processes, ABC transporter pathway, virulence proteins, and shikimate kinase pathway were significantly upregulated in a 3-day biofilm, while proteins associated with sugar transporter, degradation, and stress response were downregulated. Interestingly, in a 3-day biofilm, we observed numerous proteins involved in the central metabolism pathways which could lead to biofilm growth under diverse environments by providing an alternative metabolic route to utilize energy. In 12-day biofilms, proteins associated with peptidoglycan biosynthesis, sugar transporters, and stress responses were upregulated, whereas proteins associated with ABC transporters, DNA replication, and adhesion proteins were downregulated. Gene Ontology analysis revealed that more proteins are involved in metabolic processes in 3dwb compared with 12dwb. Furthermore, we observed significant variations in the formation of biofilms resulting from changes in the level of metabolic activity in the different growth modes of biofilms that could be a significant factor in biofilm maturation and persistence. Collectively, potential marker proteins were identified and further characterized to understand their exact role in biofilm development, which may shed light on possible new therapeutic regimes in the treatment of biofilm-related implant-associated infections.
是一种臭名昭著的生物膜产生病原体，经常从植入式医疗器械感染中分离出来。随着生物膜的老化，它对抗菌治疗的耐受性更高，导致治疗失败，并需要花费昂贵的代价来去除受感染的设备。在这项研究中，我们进行了溶液内消化，然后进行了基于 TMT 的高通量质谱分析，并研究了与 24 小时浮游生物相比，在 3 天和 12 天的生物膜中蛋白质组发生了哪些变化。结果表明，与生物合成过程、ABC 转运体途径、毒力蛋白和莽草酸激酶途径相关的蛋白质在 3 天生物膜中显著上调，而与糖转运体、降解和应激反应相关的蛋白质下调。有趣的是，在 3 天的生物膜中，我们观察到许多与中心代谢途径相关的蛋白质，这些蛋白质可以通过提供替代代谢途径来利用能量，从而在不同的环境中促进生物膜的生长。在 12 天的生物膜中，与肽聚糖生物合成、糖转运体和应激反应相关的蛋白质上调，而与 ABC 转运体、DNA 复制和粘附蛋白相关的蛋白质下调。GO 分析显示，与 12dwb 相比，3dwb 中更多的蛋白质参与代谢过程。此外，我们观察到由于生物膜不同生长模式下代谢活性水平的变化，生物膜的形成发生了显著变化，这可能是生物膜成熟和持续存在的一个重要因素。总之，鉴定了潜在的标记蛋白，并进一步对其进行了特征描述，以了解它们在生物膜发育中的确切作用，这可能为生物膜相关植入物相关感染的治疗提供新的治疗方案。
