Bicistronic Vector Expression of Recombinant Jararhagin-C and Its Effects on Endothelial Cells

• Published in: Toxins 2024-12, Vol.16 (12), p.524
• Main Authors: Ferraz, Karla Fernanda, De Lucca Caetano, Lhiri Hanna, Orefice, Daniele Pereira, Calabria, Paula Andreia Lucas, Della-Casa, Maisa Splendore, Freitas-de-Sousa, Luciana Aparecida, Beraldo-Neto, Emidio, Sanabani, Sabri Saeed, Magalhães, Geraldo Santana, Clissa, Patricia Bianca
• Format: Article
• Language: English
• Subjects: Analysis; Angiogenesis; Animals; Antibodies; Biological activity; Biological effects; Bothrops; Bothrops jararaca; Bothrops jararaca Venom - genetics; Cell adhesion & migration; Cell migration; Cell Movement - drug effects; Collagen; Crotalid Venoms - genetics; Crotalid Venoms - toxicity; Cysteine; Disintegrins - genetics; Disintegrins - pharmacology; E coli; Endothelial cells; Endothelium; Extracellular matrix; Genetic Vectors; Health aspects; Human Umbilical Vein Endothelial Cells - drug effects; Humans; Jararhagin-C; Protease; Proteases; Proteins; recombinant protein; Recombinant Proteins - genetics; Recombinant Proteins - pharmacology; snake venom disintegrins; Snakes; Toxicity; Toxins; Ubiquitin; Venom; Viral antibodies; Wound healing
• ISSN: 2072-6651; 2072-6651
• DOI: 10.3390/toxins16120524

Jararhagin-C (JarC) is a protein from the venom of 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.