Leptolysin, a Leptospira secreted metalloprotease of the pappalysin family with broad-spectrum activity
Extracellular proteolytic enzymes are produced by a variety of pathogenic microorganisms, and contribute to host colonization by modulating virulence. Here, we present a first characterization of leptolysin, a Leptospira metalloprotease of the pappalysin family identified in a previous exoproteomic...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-08-01
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| Series: | Frontiers in Cellular and Infection Microbiology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2022.966370/full |
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| author | Daniella dos Santos Courrol Cristiane Castilho Fernandes da Silva Luan Gavião Prado Luan Gavião Prado Rosa Maria Chura-Chambi Rosa Maria Chura-Chambi Ligia Morganti Gisele Oliveira de Souza Marcos Bryan Heinemann Lourdes Isaac Fernando Paiva Conte Fernanda Calheta Vieira Portaro Rodrigo Nunes Rodrigues-da-Silva Angela Silva Barbosa |
| author_facet | Daniella dos Santos Courrol Cristiane Castilho Fernandes da Silva Luan Gavião Prado Luan Gavião Prado Rosa Maria Chura-Chambi Rosa Maria Chura-Chambi Ligia Morganti Gisele Oliveira de Souza Marcos Bryan Heinemann Lourdes Isaac Fernando Paiva Conte Fernanda Calheta Vieira Portaro Rodrigo Nunes Rodrigues-da-Silva Angela Silva Barbosa |
| author_sort | Daniella dos Santos Courrol |
| collection | DOAJ |
| description | Extracellular proteolytic enzymes are produced by a variety of pathogenic microorganisms, and contribute to host colonization by modulating virulence. Here, we present a first characterization of leptolysin, a Leptospira metalloprotease of the pappalysin family identified in a previous exoproteomic study. Comparative molecular analysis of leptolysin with two other pappalysins from prokaryotes, ulilysin and mirolysin, reveals similarities regarding calcium, zinc, and arginine -binding sites conservation within the catalytic domain, but also discloses peculiarities. Variations observed in the primary and tertiary structures may reflect differences in primary specificities. Purified recombinant leptolysin of L. interrogans was obtained as a ~50 kDa protein. The protease exhibited maximal activity at pH 8.0 and 37°C, and hydrolytic activity was observed in the presence of different salts with maximum efficiency in NaCl. Substrate specificity was assessed using a small number of FRET peptides, and showed a marked preference for arginine residues at the P1 position. L. interrogans leptolysin proteolytic activity on proteinaceous substrates such as proteoglycans and plasma fibronectin was also evaluated. All proteins tested were efficiently degraded over time, confirming the protease´s broad-spectrum activity in vitro. In addition, leptolysin induced morphological alterations on HK-2 cells, which may be partially attributed to extracellular matrix (ECM) degradation. Hemorrhagic foci were observed in the dorsal skin of mice intradermally injected with leptolysin, as a plausible consequence of ECM disarray and vascular endothelium glycocalyx damage. Assuming that leptospiral proteases play an important role in all stages of the infectious process, characterizing their functional properties, substrates and mechanisms of action is of great importance for therapeutic purposes. |
| first_indexed | 2024-04-13T18:27:23Z |
| format | Article |
| id | doaj.art-5f3e9449c46e49a3a9fcf9065da85cae |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-04-13T18:27:23Z |
| publishDate | 2022-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-5f3e9449c46e49a3a9fcf9065da85cae2022-12-22T02:35:12ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882022-08-011210.3389/fcimb.2022.966370966370Leptolysin, a Leptospira secreted metalloprotease of the pappalysin family with broad-spectrum activityDaniella dos Santos Courrol0Cristiane Castilho Fernandes da Silva1Luan Gavião Prado2Luan Gavião Prado3Rosa Maria Chura-Chambi4Rosa Maria Chura-Chambi5Ligia Morganti6Gisele Oliveira de Souza7Marcos Bryan Heinemann8Lourdes Isaac9Fernando Paiva Conte10Fernanda Calheta Vieira Portaro11Rodrigo Nunes Rodrigues-da-Silva12Angela Silva Barbosa13Laboratory of Bacteriology, Butantan Institute, São Paulo, BrazilLaboratory of Structure and Function of Biomolecules, Butantan Institute, São Paulo, BrazilLaboratory of Bacteriology, Butantan Institute, São Paulo, BrazilDepartment of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, BrazilLaboratory of Bacteriology, Butantan Institute, São Paulo, BrazilCenter of Biotechnology, Energy and Nuclear Research Institute (IPEN)-CNEN/SP), São Paulo, BrazilCenter of Biotechnology, Energy and Nuclear Research Institute (IPEN)-CNEN/SP), São Paulo, BrazilDepartment of Preventive Veterinary Medicine and Animal Health, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, BrazilDepartment of Preventive Veterinary Medicine and Animal Health, School of Veterinary Medicine and Animal Science, University of São Paulo, São Paulo, BrazilDepartment of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, BrazilPilot Plant Implementation Project, Immunobiological Technology Institute, Oswaldo Cruz Foundation, Rio de Janeiro, BrazilLaboratory of Structure and Function of Biomolecules, Butantan Institute, São Paulo, BrazilLaboratory of Immunological Technology, Immunobiological Technology Institute, Oswaldo Cruz Foundation, Rio de Janeiro, BrazilLaboratory of Bacteriology, Butantan Institute, São Paulo, BrazilExtracellular proteolytic enzymes are produced by a variety of pathogenic microorganisms, and contribute to host colonization by modulating virulence. Here, we present a first characterization of leptolysin, a Leptospira metalloprotease of the pappalysin family identified in a previous exoproteomic study. Comparative molecular analysis of leptolysin with two other pappalysins from prokaryotes, ulilysin and mirolysin, reveals similarities regarding calcium, zinc, and arginine -binding sites conservation within the catalytic domain, but also discloses peculiarities. Variations observed in the primary and tertiary structures may reflect differences in primary specificities. Purified recombinant leptolysin of L. interrogans was obtained as a ~50 kDa protein. The protease exhibited maximal activity at pH 8.0 and 37°C, and hydrolytic activity was observed in the presence of different salts with maximum efficiency in NaCl. Substrate specificity was assessed using a small number of FRET peptides, and showed a marked preference for arginine residues at the P1 position. L. interrogans leptolysin proteolytic activity on proteinaceous substrates such as proteoglycans and plasma fibronectin was also evaluated. All proteins tested were efficiently degraded over time, confirming the protease´s broad-spectrum activity in vitro. In addition, leptolysin induced morphological alterations on HK-2 cells, which may be partially attributed to extracellular matrix (ECM) degradation. Hemorrhagic foci were observed in the dorsal skin of mice intradermally injected with leptolysin, as a plausible consequence of ECM disarray and vascular endothelium glycocalyx damage. Assuming that leptospiral proteases play an important role in all stages of the infectious process, characterizing their functional properties, substrates and mechanisms of action is of great importance for therapeutic purposes.https://www.frontiersin.org/articles/10.3389/fcimb.2022.966370/fullleptolysinpappalysin-1 domain proteinLeptospiraproteolytic activityextracellular matrix degradation |
| spellingShingle | Daniella dos Santos Courrol Cristiane Castilho Fernandes da Silva Luan Gavião Prado Luan Gavião Prado Rosa Maria Chura-Chambi Rosa Maria Chura-Chambi Ligia Morganti Gisele Oliveira de Souza Marcos Bryan Heinemann Lourdes Isaac Fernando Paiva Conte Fernanda Calheta Vieira Portaro Rodrigo Nunes Rodrigues-da-Silva Angela Silva Barbosa Leptolysin, a Leptospira secreted metalloprotease of the pappalysin family with broad-spectrum activity Frontiers in Cellular and Infection Microbiology leptolysin pappalysin-1 domain protein Leptospira proteolytic activity extracellular matrix degradation |
| title | Leptolysin, a Leptospira secreted metalloprotease of the pappalysin family with broad-spectrum activity |
| title_full | Leptolysin, a Leptospira secreted metalloprotease of the pappalysin family with broad-spectrum activity |
| title_fullStr | Leptolysin, a Leptospira secreted metalloprotease of the pappalysin family with broad-spectrum activity |
| title_full_unstemmed | Leptolysin, a Leptospira secreted metalloprotease of the pappalysin family with broad-spectrum activity |
| title_short | Leptolysin, a Leptospira secreted metalloprotease of the pappalysin family with broad-spectrum activity |
| title_sort | leptolysin a leptospira secreted metalloprotease of the pappalysin family with broad spectrum activity |
| topic | leptolysin pappalysin-1 domain protein Leptospira proteolytic activity extracellular matrix degradation |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2022.966370/full |
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