Data_Sheet_1_Surfaceome and Exoproteome Dynamics in Dual-Species Pseudomonas aeruginosa and Staphylococcus aureus Biofilms.xlsx

Bacterial biofilms are an important underlying cause for chronic infections. By switching into the biofilm state, bacteria can evade host defenses and withstand antibiotic chemotherapy. Despite the fact that biofilms at clinical and environmental settings are mostly composed of multiple microbial species, biofilm research has largely been focused on single-species biofilms. In this study, we investigated the interaction between two clinically relevant bacterial pathogens (Staphylococcus aureus and Pseudomonas aeruginosa) by label-free quantitative proteomics focusing on proteins associated with the bacterial cell surfaces (surfaceome) and proteins exported/released to the extracellular space (exoproteome). The changes observed in the surfaceome and exoproteome of P. aeruginosa pointed toward higher motility and lower pigment production when co-cultured with S. aureus. In S. aureus, lower abundances of proteins related to cell wall biosynthesis and cell division, suggesting increased persistence, were observed in the dual-species biofilm. Complementary phenotypic analyses confirmed the higher motility and the lower pigment production in P. aeruginosa when co-cultured with S. aureus. Higher antimicrobial tolerance associated with the co-culture setting was additionally observed in both species. To the best of our knowledge, this study is among the first systematic explorations providing insights into the dynamics of both the surfaceome and exoproteome of S. aureus and P. aeruginosa dual-species biofilms.

细菌生物被膜（bacterial biofilms）是慢性感染的重要潜在诱因。细菌通过转变为生物被膜状态，能够逃逸宿主免疫防御并耐受抗生素化疗。尽管临床与环境中的生物被膜大多由多种微生物群落构成，但现有生物被膜研究大多仍聚焦于单菌种生物被膜。本研究采用无标记定量蛋白质组学（label-free quantitative proteomics），针对两种临床相关病原菌——金黄色葡萄球菌（Staphylococcus aureus）与铜绿假单胞菌（Pseudomonas aeruginosa）展开相互作用研究，重点关注细菌细胞表面相关蛋白（表面组，surfaceome）以及被转运/释放至胞外空间的蛋白（分泌蛋白质组，exoproteome）。当与金黄色葡萄球菌共培养时，铜绿假单胞菌的表面组与分泌蛋白质组的变化显示其运动性增强、色素生成减少。在双菌种生物被膜中，我们观察到金黄色葡萄球菌内与细胞壁生物合成及细胞分裂相关的蛋白丰度降低，这提示其持久性增强。互补表型分析证实了共培养条件下铜绿假单胞菌运动性增强、色素生成减少的现象。此外，两种菌种在共培养环境下均表现出更高的抗菌药物耐受性。据我们所知，本研究是首批系统性探究金黄色葡萄球菌与铜绿假单胞菌双菌种生物被膜的表面组及分泌蛋白质组动态变化的研究之一。