2024年 新着論文 49 蛋白質発現分野から論文が発表されました

Synthesis of secretory leukocyte protease inhibitor using cell-free protein synthesis system

Odontology. 2024 Oct;112(4):1103-1112. doi: 10.1007/s10266-024-00910-8. Epub 2024 Mar 19.

Authors

Yuka Hiroshima  1 Rie Kido  2 Jun-Ichi Kido  2 Mika Bando  2 Kaya Yoshida  3 Akikazu Murakami  4 Yasuo Shinohara  5

Affiliations

  • 1 Department of Oral Microbiology, Tokushima University Graduate School of Biomedical Sciences, 3-18-15, Kuramoto, Tokushima, 770-8504, Japan. yuka.hiroshima@tokushima-u.ac.jp.
  • 2 Department of Periodontology and Endodontology, Tokushima University Graduate School of Biomedical Sciences, Tokushima, Japan.
  • 3 Department of Oral Healthcare Promotion, Tokushima University Graduate School of Biomedical Sciences, Tokushima, Japan.
  • 4 Department of Oral Microbiology, Tokushima University Graduate School of Biomedical Sciences, 3-18-15, Kuramoto, Tokushima, 770-8504, Japan.
  • 5 Institute of Advanced Medical Sciences, Tokushima University, Tokushima, Japan.

Abstract

Secretory leukocyte protease inhibitor (SLPI) functions as a protease inhibitor that modulates excessive proteolysis in the body, exhibits broad-spectrum antimicrobial activity, regulates inflammatory responses, and plays an important role in the innate immunity. The purpose of the study was to artificially synthesize a SLPI, an antimicrobial peptide, and investigate its effect on antimicrobial activity against Porphyromonas gingivalis and interleukin-6 (IL-6) production. SLPI protein with a molecular weight of approximately 13 kDa was artificially synthesized using a cell-free protein synthesis (CFPS) system and investigated by western blotting and enzyme-linked immunosorbent assay (ELISA). Disulfide bond isomerase in the protein synthesis mixture increased the amount of SLPI synthesized. The synthesized SLPI (sSLPI) protein was purified and its antimicrobial activity was investigated based on the growth of Porphyromonas gingivalis and bacterial adhesion to oral epithelial cells. The effect of sSLPI on IL-6 production in human periodontal ligament fibroblasts (HPLFs) was examined by ELISA. Our results showed that sSLPI significantly inhibited the growth of Porphyromonas gingivalis and bacterial adhesion to oral epithelial cells and further inhibited IL-6 production by HPLFs. These results suggested that SLPI artificially synthesized using the CFPS system may play a role in the prevention of periodontal diseases through its antimicrobial and anti-inflammatory effects.

Keywords: Porphyromonas gingivalis; Antimicrobial activity; Cell-free protein synthesis system; Interleukin-6; Secretory leukocyte protease inhibitor.

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