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Korean Society for Biochemistry and Molecular Biology (생화학분자생물학회)
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Structural stability for surface display of antigen 43 and application to bacterial outer membrane vesicles production

  • Gna Ahn (Department of Microbiology, Chungbuk National University) ;
  • Hyo-Won Yoon (Department of Microbiology, Chungbuk National University) ;
  • Jae-Won Choi (Department of Biopharmaceutical Sciences, Cheongju University) ;
  • Woo-Ri Shin (Department of Microbiology, Chungbuk National University) ;
  • Jiho Min (Graduate School of Semiconductor and Chemical Engineering, Jeonbuk National University) ;
  • Yang-Hoon Kim (Department of Microbiology, Chungbuk National University) ;
  • Ji-Young Ahn (Department of Microbiology, Chungbuk National University)
  • Received : 2024.04.15
  • Accepted : 2024.05.10
  • Published : 2024.08.31
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Abstract

Antigen 43 (Ag43) proteins, found on the outer membrane of Escherichia coli, are β-sheets that fold into a unique cylindrical structure known as a β-barrel. There are several known structural similarities between bacterial Ag43 autotransporters and physical components; however, the factors that stabilize the barrel and the mechanism for Ag43 passenger domain-mediated translocation across the pore of the β-barrel remain unclear. In this study, we analyzed Ag43β-enhanced green fluorescent protein chimeric variants to provide new insights into the autotransporter Ag43β-barrel assembly, focusing on the impact of the α-helical linker domain. Among the chimeric variants, Ag43β700 showed the highest surface display, which was confirmed through extracellular protease digestion, flow cytometry, and an evaluation of outer membrane vesicles (OMVs). The Ag43β700 module offered reliable information on stable barrel folding and chimera expression at the exterior of the OMVs.

Keywords

Acknowledgement

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education (No. 2020R1A6A1A06046235), the Korea Institute of Planning and Evaluation for Technology in Food, Agriculture and Forestry (IPET) through the Crop Viruses and Pests Response Industry Technology Development Program, funded by the Ministry of Agriculture, Food and Rural Affairs (MAFRA; No. 321108-04), and the Commercializations Promotion Agency for R&D Outcomes (COMPA) grant funded by the Korea government (MSIT; No. 1711173792).

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