Bicistronic Vector Expression of Recombinant Jararhagin-C and Its Effects on Endothelial Cells
Jararhagin-C (JarC) is a protein from the venom of <i>Bothrops jararaca</i> consisting of disintegrin-like and cysteine-rich domains. JarC shows a modulating effect on angiogenesis and remodeling of extracellular matrix constituents, improving wound healing in a mouse experimental model....
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MDPI AG
2024-12-01
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| Series: | Toxins |
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| Online Access: | https://www.mdpi.com/2072-6651/16/12/524 |
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| author | Karla Fernanda Ferraz Lhiri Hanna De Lucca Caetano Daniele Pereira Orefice Paula Andreia Lucas Calabria Maisa Splendore Della-Casa Luciana Aparecida Freitas-de-Sousa Emidio Beraldo-Neto Sabri Saeed Sanabani Geraldo Santana Magalhães Patricia Bianca Clissa |
| author_facet | Karla Fernanda Ferraz Lhiri Hanna De Lucca Caetano Daniele Pereira Orefice Paula Andreia Lucas Calabria Maisa Splendore Della-Casa Luciana Aparecida Freitas-de-Sousa Emidio Beraldo-Neto Sabri Saeed Sanabani Geraldo Santana Magalhães Patricia Bianca Clissa |
| author_sort | Karla Fernanda Ferraz |
| collection | DOAJ |
| description | Jararhagin-C (JarC) is a protein from the venom of <i>Bothrops jararaca</i> consisting of disintegrin-like and cysteine-rich domains. JarC shows a modulating effect on angiogenesis and remodeling of extracellular matrix constituents, improving wound healing in a mouse experimental model. JarC is purified from crude venom, and the yield is less than 1%. The aim of this work was to obtain the recombinant form of JarC and to test its biological activity. For this purpose, the bicistronic vector pSUMOUlp1 was used. This vector allowed the expression of the recombinant toxin JarC (rJarC) in fusion with the small ubiquitin-related modifier (SUMO) as well as the SUMO protease Ulp1. After expression, this protease was able to efficiently remove SUMO from rJarC inside the bacteria. rJarC free from SUMO was purified at the expected molecular mass and recognized by polyclonal anti-jararhagin antibodies. In terms of biological activity, both the native and recombinant forms showed no toxicity to the HUVEC cell line CRL1730 and were effective in modulating cell migration activity in the experimental in vitro model. These results demonstrate the successful production of rJarC and the preservation of its biological activity, which may facilitate further investigations into the therapeutic potential of this snake venom-derived protein. |
| format | Article |
| id | doaj-art-7b81d2c4304a4fffb7d902518f29f30f |
| institution | Kabale University |
| issn | 2072-6651 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Toxins |
| spelling | doaj-art-7b81d2c4304a4fffb7d902518f29f30f2024-12-27T14:56:56ZengMDPI AGToxins2072-66512024-12-01161252410.3390/toxins16120524Bicistronic Vector Expression of Recombinant Jararhagin-C and Its Effects on Endothelial CellsKarla Fernanda Ferraz0Lhiri Hanna De Lucca Caetano1Daniele Pereira Orefice2Paula Andreia Lucas Calabria3Maisa Splendore Della-Casa4Luciana Aparecida Freitas-de-Sousa5Emidio Beraldo-Neto6Sabri Saeed Sanabani7Geraldo Santana Magalhães8Patricia Bianca Clissa9Immunopathology Laboratory, Butantan Institute, São Paulo 05585-090, BrazilImmunopathology Laboratory, Butantan Institute, São Paulo 05585-090, BrazilImmunopathology Laboratory, Butantan Institute, São Paulo 05585-090, BrazilImmunopathology Laboratory, Butantan Institute, São Paulo 05585-090, BrazilImmunopathology Laboratory, Butantan Institute, São Paulo 05585-090, BrazilImmunopathology Laboratory, Butantan Institute, São Paulo 05585-090, BrazilBiochemistry Laboratory, Butantan Institute, São Paulo 05503-900, BrazilLaboratory of Medical Investigation LIM-56, Division of Dermatology, Medical School, University of São Paulo, Sao Paulo 05508-220, BrazilImmunopathology Laboratory, Butantan Institute, São Paulo 05585-090, BrazilImmunopathology Laboratory, Butantan Institute, São Paulo 05585-090, BrazilJararhagin-C (JarC) is a protein from the venom of <i>Bothrops jararaca</i> consisting of disintegrin-like and cysteine-rich domains. JarC shows a modulating effect on angiogenesis and remodeling of extracellular matrix constituents, improving wound healing in a mouse experimental model. JarC is purified from crude venom, and the yield is less than 1%. The aim of this work was to obtain the recombinant form of JarC and to test its biological activity. For this purpose, the bicistronic vector pSUMOUlp1 was used. This vector allowed the expression of the recombinant toxin JarC (rJarC) in fusion with the small ubiquitin-related modifier (SUMO) as well as the SUMO protease Ulp1. After expression, this protease was able to efficiently remove SUMO from rJarC inside the bacteria. rJarC free from SUMO was purified at the expected molecular mass and recognized by polyclonal anti-jararhagin antibodies. In terms of biological activity, both the native and recombinant forms showed no toxicity to the HUVEC cell line CRL1730 and were effective in modulating cell migration activity in the experimental in vitro model. These results demonstrate the successful production of rJarC and the preservation of its biological activity, which may facilitate further investigations into the therapeutic potential of this snake venom-derived protein.https://www.mdpi.com/2072-6651/16/12/524snake venom disintegrinsrecombinant proteinJararhagin-Cendothelial cells |
| spellingShingle | Karla Fernanda Ferraz Lhiri Hanna De Lucca Caetano Daniele Pereira Orefice Paula Andreia Lucas Calabria Maisa Splendore Della-Casa Luciana Aparecida Freitas-de-Sousa Emidio Beraldo-Neto Sabri Saeed Sanabani Geraldo Santana Magalhães Patricia Bianca Clissa Bicistronic Vector Expression of Recombinant Jararhagin-C and Its Effects on Endothelial Cells Toxins snake venom disintegrins recombinant protein Jararhagin-C endothelial cells |
| title | Bicistronic Vector Expression of Recombinant Jararhagin-C and Its Effects on Endothelial Cells |
| title_full | Bicistronic Vector Expression of Recombinant Jararhagin-C and Its Effects on Endothelial Cells |
| title_fullStr | Bicistronic Vector Expression of Recombinant Jararhagin-C and Its Effects on Endothelial Cells |
| title_full_unstemmed | Bicistronic Vector Expression of Recombinant Jararhagin-C and Its Effects on Endothelial Cells |
| title_short | Bicistronic Vector Expression of Recombinant Jararhagin-C and Its Effects on Endothelial Cells |
| title_sort | bicistronic vector expression of recombinant jararhagin c and its effects on endothelial cells |
| topic | snake venom disintegrins recombinant protein Jararhagin-C endothelial cells |
| url | https://www.mdpi.com/2072-6651/16/12/524 |
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