Crotamine: Function Diversity and Potential Applications

Marinovic, Marcelo Paradiso; Mas, Caroline Dal; Monte, Gabriela Guilherme; Felix, Daniel A.; Campeiro, Joana D’Arc; Hayashi, Mirian A. F.

doi:10.1007/978-94-007-6410-1_28

Cited by 4 publications

(5 citation statements)

References 81 publications

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“…The alignment with the sequence obtained by MALDI-ISD ( Figure 2 C) allowed us to deduce that six out of the seven unidentified residues (X) corresponded to cysteines that cannot be detected because of their engagement in disulfide bonds. The seventh unidentified residue (X38) is very likely a Lysine as this residue is conserved in all published sequences of the crotamine family [ 15 , 16 ]. Moreover, the calculated molecular mass of the sequence with K38 matches the experimental mass perfectly ( Figure 3 ).…”

Section: Resultsmentioning

confidence: 99%

Myotoxin-3 from the Pacific Rattlesnake Crotalus oreganus oreganus Venom Is a New Microtubule-Targeting Agent

et al. 2022

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Microtubule targeting agents (MTA) are anti-cancer molecules that bind tubulin and interfere with the microtubule functions, eventually leading to cell death. In the present study, we used an in vitro microtubule polymerization assay to screen several venom families for the presence of anti-microtubule activity. We isolated myotoxin-3, a peptide of the crotamine family, and three isoforms from the venom of the Northern Pacific rattlesnake Crotalus oreganus oreganus, which was able to increase tubulin polymerization. Myotoxin-3 turned out to be a cell-penetrating peptide that slightly diminished the viability of U87 glioblastoma and MCF7 breast carcinoma cells. Myotoxin 3 also induced remodeling of the U87 microtubule network and decreased MCF-7 microtubule dynamic instability. These effects are likely due to direct interaction with tubulin. Indeed, we showed that myotoxin-3 binds to tubulin heterodimer with a Kd of 5.3 µM and stoichiometry of two molecules of peptide per tubulin dimer. Our results demonstrate that exogenous peptides are good candidates for developing new MTA and highlight the richness of venoms as a source of pharmacologically active molecules.

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Section: Resultsmentioning

confidence: 99%

Myotoxin-3 from the Pacific Rattlesnake Crotalus oreganus oreganus Venom Is a New Microtubule-Targeting Agent

et al. 2022

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“…a basic characteristic and cysteine make up three precisely arranged disulphide bonds (Nicastro et al, 2003;Fadel et al, 2005). In addition, the presence of two histidine residues could provide an extra positive charge to these molecules in physiological pH and, especially, in the pH of the medium (Marinovic et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

“…Nevertheless, linear peptides usually have lower stability due to higher liability to proteases when compared with more compact disulphide-bond-stabilized structures, as for native crotamine (Marinovic et al, 2016). Despite this concern, it was verified that the DNA-dLCr complex was highly stable in embryo culture medium and any degradation in peptide-DNA was observed even after incubation for 6 h (61.3% at 360 min vs 60.5% at 0 min).…”

Section: Discussionmentioning

confidence: 99%

Disulphide-less crotamine is effective for formation of DNA–peptide complex but is unable to improve bovine embryo transfection

Freitas

Campelo

Silva

et al. 2019

Zygote

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SummaryThis study aimed to investigate the ability of disulphide-less crotamine (dLCr) to complex DNA and to evaluate whether the DNA–dLCr complex is capable of improving transfection in bovine embryos. Three experiments were performed to: (i) evaluate the formation and stability of the DNA–dLCr complex; (ii) assess the dLCr embryotoxicity by exposure of bovine embryos to dLCr; and (iii) assess the efficiency of bovine embryo transfection after microinjection of the DNA–dLCr complex or green fluorescent protein (GFP) plasmid alone (control). DNA complexation by dLCr after 30 min of incubation at 1:100 and 1:50 proportions presented higher efficiency (P < 0.05) than the two controls: native crotamine (NCr) 1:10 and lipofectamine. There was no difference between DNA–dLCr 1:25 and the controls. The DNA–dLCr complexation was evaluated at different proportions and times. In all, at least half of maximum complexation was achieved within the initial 30 min. No embryotoxicity of dLCr was verified after exposure of in vitro fertilized embryos to different concentrations of the peptide. The effectiveness of dLCr to improve exogenous gene expression was evaluated by microinjection of the DNA–dLCr complex into in vitro fertilized zygotes, followed by verification of both embryo development and GFP expression. From embryos microinjected with DNA only, 4.6% and 2.8% expressed the GFP transgene at day 5 and day 7, respectively. The DNA–dLCr complex did not increase the number of GFP-positive embryos. In conclusion, dLCr forms a complex with DNA and its application in in vitro culture is possible. However, the dLCr peptide sequence should be redesigned to improve GFP expression.

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“…In previous reports, it has been found that some of the toxin families found in the C. m. molossus venom, like SVMPs and PLA 2 s, have cytotoxic effects in different tumoral cell lines (Calderon et al, 2014;Tang et al, 2004;Boldrini-França et al, 2020;Marinovic et al, 2017;Du & Clemetson, 2002;Rivas-Mercado & Garza-Ocañas, 2017). A P-III snake venom metalloproteinase (SVMP) was identified in band A figure 1; this band may contain another protein.…”

Section: Venom Characterizationmentioning

confidence: 93%

“…Some of these protein families are responsible for many of the clinical symptoms developed by snakebite envenomation, such as local or systemic hemorrhage, neurotoxicity, and blood clotting anomalies (Masuda et al, 1998;Torii et al, 1997;Suhr & Kim, 1996;Meléndez-Martínez et al, 2017;Chellapandi, 2014;Park et al, 2009;Bénard-Valle et al, 2014;Calderon et al, 2014;Calvete et al, 2009). Interestingly enough, several studies have reported that some of these toxin families have cytotoxic action against tumoral cells (Calderon et al, 2014;Li et al, 2018b;Hayashi et al, 2012;Kerkis et al, 2014;Lee et al, 2016;Marinovic et al, 2017;Azevedo et al, 2016). Given the biological mechanisms by which they interact with different tissues, cells, and receptors, snake venom toxins can induce apoptosis, inhibit angiogenesis, tumoral growth, and cell migration (Biswas et al, 2012;Calderon et al, 2014;Al-Sadoon et al, 2013;Badr et al, 2013).…”

Section: Introductionmentioning

confidence: 99%

Cytotoxic Activity of a Northern Black-tailed Rattlesnake (Crotalus molossus molossus) Venom-Loaded in Chitosan Nanoparticles as a Potential Antitumoral System

et al. 2022

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Nanomedicine has led to the development of new materials able to improve the pharmaceutical effect of bioactive components, broadening the options of treatment for several diseases, including cancer. Chitosan (Cs) has been firmly established as biocompatible and biodegradable low-toxic polymer able to form complexes with bioactive agents, making them promising drug delivery vehicles. Additionally, some snake venom toxins such as A2 phospholipases (PLA2s), serine proteinases (SVSPs) and metalloproteinases (SVMPs) have been reported to present cytotoxic activity in different tumor cell-lines, making them an auspicious option to be used as cancer pharmaceuticals. In the present study, we identified the major proteins in a northern black-tailed rattlesnake (Crotalus molossus molossus) venom, and hemocompatibility and cytotoxic activity against T-47D breast carcinoma cells were evaluated. Afterwards, the venom was loaded into Cs nanoparticles through the ionotropic gelation process with tripolyphosphate (TPP), obtaining particles of 415.9 ± 21.67 nm and a zeta potential of +28.3 ± 1.17 mV. The Cs-Venom complex was able to deliver the venom into the breast carcinoma cells, inhibiting their viability and inducing morphological changes in the T-47D cells. Although more studies are required, we suggest the potential use of C. m. molossus venom toxins entrapped within polymer nanoparticles for the future development and research of cancer pharmaceuticals.

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Crotamine: Function Diversity and Potential Applications

Cited by 4 publications

References 81 publications

Myotoxin-3 from the Pacific Rattlesnake Crotalus oreganus oreganus Venom Is a New Microtubule-Targeting Agent

Myotoxin-3 from the Pacific Rattlesnake Crotalus oreganus oreganus Venom Is a New Microtubule-Targeting Agent

Disulphide-less crotamine is effective for formation of DNA–peptide complex but is unable to improve bovine embryo transfection

Cytotoxic Activity of a Northern Black-tailed Rattlesnake (Crotalus molossus molossus) Venom-Loaded in Chitosan Nanoparticles as a Potential Antitumoral System

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