Inhibition of Tannerpin-M encoded by periodontal pathogens on serine proteases released by granulocytes
Objective·To prepare a serine protease inhibitor (Serpin) derived from Tannerella which is associated with periodontosis, and analyze its specificity in inhibiting target proteases and its structural characteristics.Methods·Through amino acid sequence analysis, a Serpin from the human oral microbiom...
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Editorial Office of Journal of Shanghai Jiao Tong University (Medical Science)
2024-12-01
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| Series: | Shanghai Jiaotong Daxue xuebao. Yixue ban |
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| Online Access: | https://xuebao.shsmu.edu.cn/article/2024/1674-8115/1674-8115-2024-44-12-1536.shtml |
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| author | PAN Zihao XU Jiawei ZHOU Aiwu |
| author_facet | PAN Zihao XU Jiawei ZHOU Aiwu |
| author_sort | PAN Zihao |
| collection | DOAJ |
| description | Objective·To prepare a serine protease inhibitor (Serpin) derived from Tannerella which is associated with periodontosis, and analyze its specificity in inhibiting target proteases and its structural characteristics.Methods·Through amino acid sequence analysis, a Serpin from the human oral microbiome database (eHOMD) was selected and expressed in Escherichia coli. The recombinant protein was purified using methods such as nickel ion affinity chromatography. Its specificity in inhibiting serine proteases was analyzed, followed by an analysis of its three-dimensional spatial structure using structural biology methods.Results·A novel Serpin, named Tannerpin-M, with methionine as the active center P1 residue, was identified, and a high-purity recombinant protein was successfully prepared from Escherichia coli BL21 (DE3). Further activity testing demonstrated that recombinant Tannerpin-M could effectively form SDS-stable covalent complexes with proteases derived from granulocytes (human neutrophil elastase, cathepsin G, and proteinase 3), as well as with other proteases including kallikrein 1 (KLK1), KLK7, and elastase. Tannerpin-M inhibited KLK7 with a second-order association rate constant of 4.12×104 L/(mol·s). The crystal structure of Tannerpin-M in its relaxed state conformation was resolved at a resolution of 2.4 Å (1 Å=0.1 nm). It revealed that Tannerpin-M possessed a significantly elongated reactive center loop and could undergo the classical conformational transition from a stressed to a relaxed state.Conclusion·Tannerpin-M, derived from oral pathogenic bacteria, is a typical inhibitory Serpin, and can effectively inhibit the serine protease released by granulocytes, by which it may protect the oral pathogenic bacteria from attacks of the human immune system. |
| format | Article |
| id | doaj-art-b53f5f3cfcf54dc08c416f69c301e77f |
| institution | Kabale University |
| issn | 1674-8115 |
| language | zho |
| publishDate | 2024-12-01 |
| publisher | Editorial Office of Journal of Shanghai Jiao Tong University (Medical Science) |
| record_format | Article |
| series | Shanghai Jiaotong Daxue xuebao. Yixue ban |
| spelling | doaj-art-b53f5f3cfcf54dc08c416f69c301e77f2025-01-08T05:52:54ZzhoEditorial Office of Journal of Shanghai Jiao Tong University (Medical Science)Shanghai Jiaotong Daxue xuebao. Yixue ban1674-81152024-12-0144121536154410.3969/j.issn.1674-8115.2024.12.0061674-8115（2024）12-1536-09Inhibition of Tannerpin-M encoded by periodontal pathogens on serine proteases released by granulocytesPAN Zihao0XU Jiawei1ZHOU Aiwu2Department of Biopharmaceuticals and Food Science, School of Pharmacy, Nanjing University of Chinese Medicine, Nanjing210023, ChinaDepartment of Biological Engineering, Zhuhai Campus of Zunyi Medical University, Zhuhai519040, ChinaDepartment of Pathophysiology, Shanghai Jiao Tong University College of Basic Medical Sciences, Shanghai200025, ChinaObjective·To prepare a serine protease inhibitor (Serpin) derived from Tannerella which is associated with periodontosis, and analyze its specificity in inhibiting target proteases and its structural characteristics.Methods·Through amino acid sequence analysis, a Serpin from the human oral microbiome database (eHOMD) was selected and expressed in Escherichia coli. The recombinant protein was purified using methods such as nickel ion affinity chromatography. Its specificity in inhibiting serine proteases was analyzed, followed by an analysis of its three-dimensional spatial structure using structural biology methods.Results·A novel Serpin, named Tannerpin-M, with methionine as the active center P1 residue, was identified, and a high-purity recombinant protein was successfully prepared from Escherichia coli BL21 (DE3). Further activity testing demonstrated that recombinant Tannerpin-M could effectively form SDS-stable covalent complexes with proteases derived from granulocytes (human neutrophil elastase, cathepsin G, and proteinase 3), as well as with other proteases including kallikrein 1 (KLK1), KLK7, and elastase. Tannerpin-M inhibited KLK7 with a second-order association rate constant of 4.12×104 L/(mol·s). The crystal structure of Tannerpin-M in its relaxed state conformation was resolved at a resolution of 2.4 Å (1 Å=0.1 nm). It revealed that Tannerpin-M possessed a significantly elongated reactive center loop and could undergo the classical conformational transition from a stressed to a relaxed state.Conclusion·Tannerpin-M, derived from oral pathogenic bacteria, is a typical inhibitory Serpin, and can effectively inhibit the serine protease released by granulocytes, by which it may protect the oral pathogenic bacteria from attacks of the human immune system.https://xuebao.shsmu.edu.cn/article/2024/1674-8115/1674-8115-2024-44-12-1536.shtmlserine protease inhibitortannerellaprokaryotic expressioncrystal structureperiodontal pathogen |
| spellingShingle | PAN Zihao XU Jiawei ZHOU Aiwu Inhibition of Tannerpin-M encoded by periodontal pathogens on serine proteases released by granulocytes Shanghai Jiaotong Daxue xuebao. Yixue ban serine protease inhibitor tannerella prokaryotic expression crystal structure periodontal pathogen |
| title | Inhibition of Tannerpin-M encoded by periodontal pathogens on serine proteases released by granulocytes |
| title_full | Inhibition of Tannerpin-M encoded by periodontal pathogens on serine proteases released by granulocytes |
| title_fullStr | Inhibition of Tannerpin-M encoded by periodontal pathogens on serine proteases released by granulocytes |
| title_full_unstemmed | Inhibition of Tannerpin-M encoded by periodontal pathogens on serine proteases released by granulocytes |
| title_short | Inhibition of Tannerpin-M encoded by periodontal pathogens on serine proteases released by granulocytes |
| title_sort | inhibition of tannerpin m encoded by periodontal pathogens on serine proteases released by granulocytes |
| topic | serine protease inhibitor tannerella prokaryotic expression crystal structure periodontal pathogen |
| url | https://xuebao.shsmu.edu.cn/article/2024/1674-8115/1674-8115-2024-44-12-1536.shtml |
| work_keys_str_mv | AT panzihao inhibitionoftannerpinmencodedbyperiodontalpathogensonserineproteasesreleasedbygranulocytes AT xujiawei inhibitionoftannerpinmencodedbyperiodontalpathogensonserineproteasesreleasedbygranulocytes AT zhouaiwu inhibitionoftannerpinmencodedbyperiodontalpathogensonserineproteasesreleasedbygranulocytes |