| Autor: |
Ferraz KF; Immunopathology Laboratory, Butantan Institute, São Paulo 05585-090, Brazil., De Lucca Caetano LH; Immunopathology Laboratory, Butantan Institute, São Paulo 05585-090, Brazil., Orefice DP; Immunopathology Laboratory, Butantan Institute, São Paulo 05585-090, Brazil., Calabria PAL; Immunopathology Laboratory, Butantan Institute, São Paulo 05585-090, Brazil., Della-Casa MS; Immunopathology Laboratory, Butantan Institute, São Paulo 05585-090, Brazil., Freitas-de-Sousa LA; Immunopathology Laboratory, Butantan Institute, São Paulo 05585-090, Brazil., Beraldo-Neto E; Biochemistry Laboratory, Butantan Institute, São Paulo 05503-900, Brazil., Sanabani SS; Laboratory of Medical Investigation LIM-56, Division of Dermatology, Medical School, University of São Paulo, Sao Paulo 05508-220, Brazil., Magalhães GS; Immunopathology Laboratory, Butantan Institute, São Paulo 05585-090, Brazil., Clissa PB; Immunopathology Laboratory, Butantan Institute, São Paulo 05585-090, Brazil. |
| Abstrakt: |
Jararhagin-C (JarC) is a protein from the venom of Bothrops jararaca 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. |
