Crotamine-like from Southern Pacific rattlesnake (Crotalus oreganus helleri) Venom acts on human leukemia (K-562) cell lines and produces ultrastructural changes on mice adrenal gland

Abstract

Crotamine is a cationic, non-enzymatic, protein integrating a minor family of myotoxins, composed of 42 amino acid residues, described in Viperidae and Crotalidae snake's families that has been used in neuroscience research, drug progressing and molecular diversity reports. Crotamine-like protein (CLP) from C.o.helleri venom was isolated in fraction 5 from 7 peaks obtained by sulfopropyl waters protein pak cationic exchange column. In tricine-SDS-PAGE under non-reduced conditions this CLP showed a single band of ~8 kDa molecular weight. CLP induced toxicity of K-562 cells with a CC of 11.09 µM. In mice adrenal gland after 24 h of CLP injection, cortical cells exhibited swollen mitochondria with scarce tubular cristae, some elements of smooth and rough endoplasmic reticula, widened nuclear envelope, slightly osmiophilic lipid droplets, and autophagic vacuoles. In some areas cortical cells plasma membrane and endothelial walls disappeared, which indicated a necrosis process. In other areas, endothelial cell cytoplasm did not present the normal caveolae and pinocytotic vesicles. To our knowledge, this is the first report on mice adrenal gland damages, caused by the injection of CLP from rattlesnakes. Our results propose that adrenal cortex lesions may be significant in the envenoming etiopathogenesis by CLP.

A new crotamine-like from the rattlesnake (Crotalus durissus cumanensis) venom causing damages: Qualitative and Quantitative Cytotoxic Studies on subcellular and neuromuscular structures

“…In recent years, several neuro and myotoxins have been isolated from snake venoms, and the mechanism of action of their components has been investigated [2,3,8,29], since crotamine is one of the important components of Crotalidae venom that combines cytotoxic and neurotoxic properties [5], it was carried out this study of the alterations in the neuro-motor plates produced by the action of purified C-L as described in results sections.…”

“…Purification of C-L from the rattlesnake venom was carried out by one chromatographic step [29]. Crude venom (Cdc) (30 milligrams-mg by protein estimation) [33] was diluted to 1.0 millilitres (mL) of 50 milli Molar (mM) Tris-HCl buffer, pH 8.2, and then applied on a Mono S HR 10/10 column (GE Healthcare Biosciences Ltd, USA) equilibrated with an equivalent buffer.…”

“…The venoms of Crotalus snakes are responsible for pathological sub-cellular lesions, which have been described by electron microscopy techniques in several previous works [2,8,25,29]. It is common that in venoms scientific research there is a relationship between the enzymatic and clinical pathological pictures; however, when adding the sub-cellular level of description that offers a tool such as Electron Microscopy (EM), the understanding and the pathological analysis is necessarily improved.…”

Qualitative and Quantitative Ultrastructural Analysis of the Mitochondria from Adrenal Gland Cortex Under the Action of Viperidae family Snake Venoms

“…There is a difference between Crotalus and Bothrops envenomation, possibly related to the toxic mechanism present in venoms. Rattlesnake (Crotalus) venom contains a variety of peptides, enzymes and toxins such as the crotoxin, a toxin that creates an effect of intense muscular damage and bleeding blocking platelet aggregation (Hati et al, 1999) or crotamine that has cytotoxic effects (Sánchez et al, 2018). Also there are metalloproteases and phospholipases acting on capillary endothelial cells (Avila-Agüero et al, 1999;Hernández et al, 2006), which cause haemorrhages in many organs, including the adrenal glands (Yeh et al, 2001).…”

Effect of the snake venom component crotamine on lymphatic endothelial cell responses and lymph transport

“…16,19 Crotamine is a 42 amino acid long, cell penetrating peptide similar to beta defensins that was originally described as a myotoxin; however, that assertion has been challenged recently. 21,22 It is, however, an inhibitor of the K V α1.X family of voltage-gated potassium channels comprised of 8 members (K V 1.1-1.8). 23 Crotamine is commonly associated with paraplegia in animal studies due to it's effects on the nervous system.…”