cDNA Cloning, Heterologous Expression, Cytotoxicity, and Inhibitory Effects of a Disintegrin from Bothrops ammodytoides Venom

Abstract An mRNA transcript that codes for a Disintegrin and Metalloprotease from venom gland of the viper Bothrops ammodytoides was isolated. Vector pCR®2.1-TOPO used to clone then subcloned in pET-28a vector express protein E. coli strain BL21. The recombinant disintegrin, HisrDisintegrin, comprises 38 residues at N-terminal 75 Disintegrin, including 14 cysteines or 7 disulfide bonds. HisrDisintegrin obtained soluble intracellular fraction separated by affinity chromatography. experimental molecular mass 11,750.8 Da, agreed with its theoretical value, cleavage Met residue caused bacterial post-translational modification. folded vitro conditions purified, finding main comparable mass. contains an alpha/beta structure, as observed circular dichroism. Its biological activity demonstrated growth reduction human endothelial (HMEC-1) cells mammary adenocarcinoma (MDA-MB-231). Furthermore, inhibits adhesion HMEC-1 MDA-MB-231 proteins extracellular matrix (laminin (LN), fibronectin (FN), vitronectin (VN)).