Micrurus surinamensis Peruvian snake venom: Cytotoxic activity and purification of a C-type lectin protein (Ms-CTL) highly toxic to cardiomyoblast-derived H9c2 cells

Micrurus surinamensis (Cuvier, 1817), popularly known as aquatic coral snake, has a broad geographic distribution in the Rainforest of South America. The purpose of this study was to investigate the cytotoxic effect caused by M. surinamensis venom in H9c2 cardiomyoblast cells and to identify protein components involved in cardiotoxic processes. Venom cardiotoxic potential is evidenced by cell viability reduction in a concentration-dependent manner. We have purified one of venom components responsible for this effect after three chromatographic steps: a cytotoxic 23.461 kDa protein, as determined by mass spectrometry. A 19-residue sequence (DCPSGWSSYEGSCYNFFQR) of the purified protein was deduced by MS/MS and exhibited high homology with N-terminal region of C-type lectin from snake venoms. This protein was named Ms-CTL. Morphologically, H9c2 incubation with Ms-CTL led to a significant cellular retraction and formation of cellular aggregates, as observed by microscopy phase-contrast images. Our results indicate that M. surinamensis venom is highly toxic to H9c2 cardiomyoblast cell and less or not cytotoxic to other cell lines, such as HaCat, VERO and U373. Results presented herein will help understanding the mechanisms that underlie cellular damage and tissue destruction, being useful in the development of alternative therapies against these coral snake bites.