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

The Journal of Antibiotics volume 76, pages 406–415 (2023)Cite this article

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Abstract

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 MIC50/MIC90 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 12S. 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.

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Acknowledgements

The authors thank Dr. Weiguang Pan and Jie Lian (Department of Laboratory Medicine, Shenzhen Nanshan People’s Hospital and the 6th Affiliated Hospital of Shenzhen University Medical School) for helping identify and preserve the S. aureus strains.

Funding

This work was supported by the following grants: The Jiangsu Synergetic Innovation Center for Advanced Bio-Manufacture (No. XTD2210); National Natural Science Foundation of China (grants 82172283); Natural Science Foundation of Guangdong Province, China (No. 2020A1515111146, 2020A1515011049, 2020A1515010979); Shenzhen Key Medical Discipline Construction Fund (No.SZXK06162); the Science, Technology and Innovation Commission of Shenzhen Municipality basic research funds (grants JCYJ20180302144403714; JCYJ20220530141810023, JCYJ20220530142006015) and key funds JCYJ20180508162403996).

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  1. These authors contributed equally: Yue Luo, Zewen Wen, Yanpeng Xiong

Authors and Affiliations

  1. College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, 211816, China

    Yue Luo, Yanpeng Xiong, Xuecheng Chen, Yalan Peng & Shiqing Han

  2. Department of Infectious Diseases and Shenzhen Key Laboratory for Endogenous Infections, Shenzhen Nanshan People’s Hospital and the 6th Affiliated Hospital of Shenzhen University Medical School, Shenzhen, 518052, China

    Zewen Wen, Zonglin Shen, Peiyu Li, Qiwen Deng, Zhijian Yu & Jinxin Zheng

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Contributions

YL participated in the design of this study and the synthesis of bithionol biotinylated probes Bio-A2, and drafted manuscript. ZW conducted the immunoprecipitation, participated in the measurement of the growth of S. aureus planktonic cells, and conducted LC-MS/MS experiments. YX participated in the synthesis of bithionol biotinylated probes Bio-A2, antimicrobial susceptibility testing and molecular docking. XC participated in the synthesis of bithionol biotinylated probes Bio-A2. ZS and PL collected S. aureus strains and conducted biofilm assay, and participated in the measurement of the growth of S. aureus planktonic cells. YP participated in the synthesis of bithionol biotinylated probes Bio-A2. QD and ZY participated in the design of this study, and participate in guiding and analyzing immunoprecipitation and LC-MS/MS experiments and data analysis, and reviewed this manuscript. JZ and SH designed the study, participated in the synthesis of bithionol biotinylated probes Bio-A2 and immunoprecipitation and LC-MS/MS experiments, conducted the data analysis, and provided critical revisions of the manuscript for valuable intellectual content.

Corresponding authors

Correspondence to Jinxin Zheng or Shiqing Han.

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The authors declare no competing interests.

Ethical approval

All procedures involving human subjects were approved by the institutional ethical committee of Shenzhen Nanshan People’s Hospital and the 6th Affiliated Hospital of Shenzhen University Medical School. Isolates were collected as part of the routine clinical management of patients, according to the national guidelines in China.

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Luo, Y., Wen, Z., Xiong, Y. et al. The potential target of bithionol against Staphylococcus aureus: design, synthesis and application of biotinylated probes Bio-A2. J Antibiot 76, 406–415 (2023). https://doi.org/10.1038/s41429-023-00618-x

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