The potential target of bithionol against Staphylococcus aureus: design, synthesis and application of biotinylated probes Bio-A2

• Published in: Journal of antibiotics 2023-07, Vol.76 (7), p.406-415
• Main Authors: Luo, Yue, Wen, Zewen, Xiong, Yanpeng, Chen, Xuecheng, Shen, Zonglin, Li, Peiyu, Peng, Yalan, Deng, Qiwen, Yu, Zhijian, Zheng, Jinxin, Han, Shiqing
• Format: Article
• Language: English
• Subjects: 631/326/22/1290; 631/92/609; 692/699/255/1318; 82; 82/58; Alanine; Anti-Bacterial Agents - chemistry; Anti-Bacterial Agents - pharmacology; Antibacterial activity; Bacteriology; Biofilms; Biomedical and Life Sciences; Bioorganic Chemistry; Bithionol; Chromatography, Liquid; Clinical isolates; DNA probes; DNA topoisomerase; Humans; Hydrogen bonding; Hydrogen bonds; Immunoprecipitation; Life Sciences; Medicinal Chemistry; Methicillin-Resistant Staphylococcus aureus; Microbial Sensitivity Tests; Microbiology; Molecular docking; Molecular Docking Simulation; Organic Chemistry; Peptides; Probes; Proteins; Staphylococcal Infections - microbiology; Staphylococcus aureus; Staphylococcus aureus - genetics; Tandem Mass Spectrometry; Translocase; tRNA Ala
• ISSN: 0021-8820; 1881-1469
• DOI: 10.1038/s41429-023-00618-x

This study aims to explore the potential targets of bithionol in Staphylococcus aureus .The four bithionol biotinylated probes Bio-A2-1 , Bio-A2-2 , Bio-A2-3 , and Bio-A2-4 were synthesized, the minimal inhibitory concentrations (MICs) of these probes against S. aureus were determined. The bithionol binding proteins in S. aureus were identified through immunoprecipitation and LC-MS/MS with bithionol biotinylated probe. The biotinylated bithionol probes Bio-A2-1 and Bio-A2-3 displayed antibacterial activities against S. aureus . The Bio-A2-1 showed lower MICs than Bio-A2-3 , and both with the MIC 50 /MIC 90 at 12.5/12.5 μM against S. aureus clinical isolates. The inhibition rates of bithionol biotinylated probes Bio-A2-1 and Bio-A2-3 on the biofilm formation of S. aureus were comparable to that of bithionol, and were stronger than that of Bio-A2-2 and Bio-A2-4 . The biofilm formation of 10 out of 12 S. aureus clinical isolates could be inhibited by Bio-A2-1 (at 1/4×, or 1/2× MICs). There are three proteins identified in S. aureus through immunoprecipitation and LC-MS/MS with bithionol biotinylated probe Bio-A2-1 : Protein translocase subunit SecA 1 ( secA1 ), Alanine--tRNA ligase ( alaS ) and DNA gyrase subunit A ( gyrA ), and in which the SecA1 protein the highest coverage and the most unique peptides. The LYS112, GLN143, ASP213, GLY496 and ASP498 of SecA1 protein act as hydrogen acceptors to form 6 hydrogen bonds with bithionol biotinylated probe Bio-A2-1 by molecular docking analysis. In conclusion, the bithionol biotinylated probe Bio-A2-1 has antibacterial and anti-biofilm activities against S. aureus , and SecA1 was probably one of the potential targets of bithionol in S. aureus .