Extraction, identification, and immunogenicity analysis of Trueperella pyogenes heparin high affinity proteins
10.13200/j.cnki.cjb.004454
- VernacularTitle:化脓隐秘杆菌肝素高亲和蛋白的提取、鉴定及其免疫原性分析
- Author:
XU Dengfeng
- Publication Type:Journal Article
- Keywords:
Trueperella pyogenes(T.pyogenes);
Heparin high affinity proteins;
Immunogenicity;
Pyolysin(PLO)
- From:
Chinese Journal of Biologicals
2025;38(04):413-417
- CountryChina
- Language:Chinese
-
Abstract:
Objective To extract and identify Trueperella pyogenes(T.pyogenes) heparin high affinity proteins(THHAPs),and analyze their immunogenicity, so as to provide a reference for exploring the interaction between T.pyogenes and heparin.Methods The surface proteins of T.pyogenes were extracted using a bacterial membrane protein extraction kit. THHAPs were extracted from the surface proteins by adsorbing with heparin-agarose, washing with 1 mol/L NaCl solution, and eluting with 8 mol/L urea solution, which were then analyzed using SDS-PAGE and liquid chromatography-tandem mass spectrometry(LC-MS/MS). Bioinformatics algorithms were used to analyze the subcellular localization, signal peptides, transmembrane domains and immunogenicity of THHAPs. Fifteen female Kunming mice were immunized with THHAPs. After two weeks of the second immunization, five mice were collected for blood samples from the orbital vein and the serum was isolated, and the other 10 mice were intraperitoneally injected with 9 × 108CFU of T.pyogenes. The immunoprotective effect and antiserum hemolytic inhibitory activity were detected.Results THHAPs were observed to form a major band accounting for 98% of the protein content. A total of 1 767 pyolysin(PLO) peptides were detected, accounting for 98. 06% of the detected peptides(1767/1 802). THHAPs were composed of 1 cell wall protein, 11 secreted proteins, 1 lipoprotein, 7 membrane proteins, and 1cytoplasmic protein, 8 proteins among which might be antigens with immunoprotective properties. The mice(9/10) immunized with THHAPs were able to resist T. pyogenes infection, and their antisera exhibited hemolytic inhibitory activity.Conclusion THHAPs have been successfully extracted from the surface proteins of T. pyogenes, with PLO as their major component, which can induce a protective immune response in mice.
