Sharma Ayushi, Bansal Saurabh, Kumari Neha, Vashistt Jitendraa, Shrivastava Rahul
Department of Biotechnology and Bioinformatics, Jaypee University of Information Technology, Waknaghat, Solan, 173234, H.P, India.
Appl Microbiol Biotechnol. 2023 Oct;107(19):6029-6046. doi: 10.1007/s00253-023-12705-y. Epub 2023 Aug 5.
Biofilm formation by Mycobacterium fortuitum causes serious threats to human health due to its increased contribution to nosocomial infections. In this study, the first comprehensive global proteome analysis of M. fortuitum was reported under planktonic and biofilm growth states. A label-free Q Exactive Quadrupole-Orbitrap tandem mass spectrometry analysis was performed on the protein lysates. The differentially abundant proteins were functionally characterized and re-annotated using Blast2GO and CELLO2GO. Comparative analysis of the proteins among two growth states provided insights into the phenotypic switch, and fundamental pathways associated with pathobiology of M. fortuitum biofilm, such as lipid biosynthesis and quorum-sensing. Interaction network generated by the STRING database revealed associations between proteins that endure M. fortuitum during biofilm growth state. Hypothetical proteins were also studied to determine their functional alliance with the biofilm phenotype. CARD, VFDB, and PATRIC analysis further showed that the proteins upregulated in M. fortuitum biofilm exhibited antibiotic resistance, pathogenesis, and virulence. Heatmap and correlation analysis provided the biomarkers associated with the planktonic and biofilm growth of M. fortuitum. Proteome data was validated by qPCR analysis. Overall, the study provides insights into previously unexplored biochemical pathways that can be targeted by novel inhibitors, either for shortened treatment duration or for eliminating biofilm of M. fortuitum and related nontuberculous mycobacterial pathogens. KEY POINTS: • Proteomic analyses of M. fortuitum reveals novel biofilm markers. • Acetyl-CoA acetyltransferase acts as the phenotype transition switch. • The study offers drug targets to combat M. fortuitum biofilm infections.
偶然分枝杆菌形成的生物膜对人类健康构成严重威胁，因为它对医院感染的影响日益增加。在本研究中，首次报道了偶然分枝杆菌在浮游和生物膜生长状态下的全面全球蛋白质组分析。对蛋白质裂解物进行了无标记的Q Exactive四极杆-轨道阱串联质谱分析。使用Blast2GO和CELLO2GO对差异丰富的蛋白质进行功能表征和重新注释。两种生长状态下蛋白质的比较分析为表型转换以及与偶然分枝杆菌生物膜病理生物学相关的基本途径（如脂质生物合成和群体感应）提供了见解。STRING数据库生成的相互作用网络揭示了在生物膜生长状态下维持偶然分枝杆菌的蛋白质之间的关联。还对假设蛋白质进行了研究，以确定它们与生物膜表型的功能联盟。CARD、VFDB和PATRIC分析进一步表明，在偶然分枝杆菌生物膜中上调的蛋白质表现出抗生素抗性、致病机制和毒力。热图和相关性分析提供了与偶然分枝杆菌浮游和生物膜生长相关的生物标志物。蛋白质组数据通过qPCR分析进行了验证。总体而言，该研究为以前未探索的生化途径提供了见解，这些途径可成为新型抑制剂的作用靶点，用于缩短治疗时间或消除偶然分枝杆菌和相关非结核分枝杆菌病原体的生物膜。关键点：•偶然分枝杆菌的蛋白质组分析揭示了新的生物膜标志物。•乙酰辅酶A乙酰转移酶作为表型转换开关。•该研究提供了对抗偶然分枝杆菌生物膜感染的药物靶点。
