Al Nahhas Dina, Pisanu Salvatore, Tanca Alessandro, Paglietti Bianca, Santona Antonella, Uzzau Sergio, Pagnozzi Daniela
Porto Conte Ricerche S.R.L., Tramariglio, Alghero, Italy.
Department of Biomedical Sciences, University of Sassari, Sassari, Italy.
Sci Rep. 2025 Aug 5;15(1):28656. doi: 10.1038/s41598-025-04224-9.
The ability of methicillin-resistant Staphylococcus aureus (MRSA) to adapt to environmental stressors is crucial for its survival and persistence. This study used shotgun proteomics to analyze the protein profiles of MRSA strains ST398 and JE2 under three experimental conditions: EC1 (control, 37 °C, pH 7), EC2 (35 °C, pH 6), and EC3 (35 °C, pH 6 with 5% NaCl). Proteins were extracted and digested with trypsin using filter-aided sample preparation (FASP), followed by LC-MS/MS analysis. Raw data were analyzed using Proteome Discoverer software with label-free quantification (LFQ), identifying an average of 2640 ± 70 and 2832 ± 154 proteins in ST398 and JE2, respectively. Qualitative and quantitative differences in protein expression were observed between strains and conditions. Under EC2, ABC transporters increased and arginine metabolism was upregulated, suggesting metabolic adaptation to acidic stress. Purine metabolism was downregulated, indicating a metabolic shift. Under EC3 condition, NaCl addition induced glycine-betaine biosynthesis and proteins involved in potassium limitation. JE2 exhibited a more pronounced oxidative and stress response under salt stress, indicating a strain-specific adaptation. These findings highlight strain-specific proteomic responses in MRSA, highlighting the roles of arginine, glycine-betaine, and purine metabolism, along with stress-response proteins, in coping with environmental stressors.
耐甲氧西林金黄色葡萄球菌(MRSA)适应环境应激源的能力对其生存和持续存在至关重要。本研究使用鸟枪法蛋白质组学分析了MRSA菌株ST398和JE2在三种实验条件下的蛋白质谱:EC1(对照,37°C,pH 7)、EC2(35°C,pH 6)和EC3(35°C,pH 6并添加5% NaCl)。使用滤膜辅助样品制备(FASP)法提取蛋白质并用胰蛋白酶消化,随后进行液相色谱-串联质谱(LC-MS/MS)分析。使用无标记定量(LFQ)的Proteome Discoverer软件分析原始数据,分别在ST398和JE2中平均鉴定出2640±70和2832±154种蛋白质。观察到菌株和条件之间蛋白质表达存在定性和定量差异。在EC2条件下,ABC转运蛋白增加,精氨酸代谢上调,表明对酸性应激的代谢适应。嘌呤代谢下调,表明代谢转变。在EC3条件下,添加NaCl诱导了甘氨酸-甜菜碱生物合成以及与钾限制相关的蛋白质。JE2在盐胁迫下表现出更明显的氧化和应激反应,表明存在菌株特异性适应。这些发现突出了MRSA中菌株特异性的蛋白质组学反应,强调了精氨酸、甘氨酸-甜菜碱和嘌呤代谢以及应激反应蛋白在应对环境应激源中的作用。