Partial purification of peptides present in the Tityus macrochirus (Buthidae) scorpion venom and preliminary assessment of their cytotoxicity

Clara Andrea Rincón-Cortés, Edgar Antonio Reyes-Montaño, Nohora Angélica Vega-Castro, .

Keywords: Scorpion venoms, peptides/isolation & purification, neoplasms

Abstract

Introduction: Scorpion venom contains peptides with neurotoxic action primarily active on ion channels in the nervous system of insects and mammals. They are also characterized as cytolytic and anticancer, biological characteristics that have not yet been reported for the Tityus macrochirus venom.
Objective: To assess if the total T. macrochirus venom and the fraction of partially purified peptides decrease the viability of various tumor-derived cell lines.
Materials and methods: The scorpion venom was collected by electrical stimulation and, subsequently, subjected to chromatography, electrophoresis, and ultrafiltration with Amicon Ultra 0.5® membranes for the partial identification and purification of its peptides. The cytotoxic activity of the venom and the peptides fraction trials on tumor-derived cell lines were carried out by the MTT method.
Results: The T. macrochirus scorpion venom has peptides with molecular weights ranging between 3 and 10 kDa. They were partially purified using the ultrafiltration technique, and assessed by the RP-HPLC method. Cytotoxicity trials with the whole T. macrochirus venom showed a higher viability decrease on the PC3 cell line compared to the other cell lines assessed, while the partially purified peptides decreased the HeLa cell line viability.
Conclusion: Peptides in the T. macrochirus scorpion venom showed cytotoxic activity on some tumorderived cell lines. We observed some degree of selectivity against other cell lines assessed.

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  • Clara Andrea Rincón-Cortés Grupo de Investigación en Proteínas, Departamento de Química, Universidad Nacional de Colombia, Bogotá, D.C., Colombia Universidad de Ciencias Aplicadas y Ambientales - U.D.C.A., Bogotá, D.C., Colombia
  • Edgar Antonio Reyes-Montaño Grupo de Investigación en Proteínas, Departamento de Química, Universidad Nacional de Colombia, Bogotá, D.C., Colombia
  • Nohora Angélica Vega-Castro Grupo de Investigación en Proteínas, Departamento de Química, Universidad Nacional de Colombia, Bogotá, D.C., Colombia

References

Possani LD, Martin BM, Svendsen JB, Rode GS, Erickson BW. Scorpion toxins from Centruroides noxius and Tityus serrulatus primary structures and sequence comparison by metric analysis. Biochem J. 1985;229:739-50. http://dx.doi.org/10.1042/bj2290739

Escobar E, Velásquez L, Rivera C. Separación e identificación de algunas toxinas del veneno de Centruroides margaritatus (Gervais, 1841) (Scorpiones: Buthidae). Rev Perú Biol. 2003;10:217-20. http://dx.doi.org/10.15381/rpb.v10i2.2507

Barona J, Batista VF, Zamudio FZ, Gómez-Lagunas F, Wanke E, Otero R, et al. Proteomic analysis of the venom and characterization of toxins specific for Na+- and K+- channels from the Colombian scorpion Tityus pachyurus. Biochim Biophys Acta. 2006;1764:76-84. http://dx.doi.org/10.1016/j.bbapap.2005.08.010

Alves RS, Ximenes RM, Antonio RC, Nascimento NR, Martins RD, Rabello MM, et al. Isolation, homology modeling and renal effects of a C-type natriuretic peptide from the venom of the Brazilian yellow scorpion (Tityus serrulatus). Toxicon. 2013;74:19-26. http://dx.doi.org/10.1016/j.toxicon.2013.07.016

Ferreira LA, Alves EW, Henriques OB. Peptide T, a novel bradykinin potentiator isolated from Tityus serrulatus scorpion venom. Toxicon. 1993;31:941-7. http://dx.doi.org/10.1016/0041-0101(93)90253-F

Dai C, Ma Y, Zhao Z, Zhao R, Wang Q, Wu Y, et al. Mucroporin, the first cationic host defense peptide from the venom of Lychas mucronatus. Antimicrob Agents Chemother. 2008;52:3967-72. http://dx.doi.org/10.1128/AAC.00542-08

Guerrero-Vargas JA, Moura CB, Quintero-Herna V, Possani LD, Schwartz EF. Identification and phylogenetic analysis of Tityus pachyrus and Tityus obscurus novel putative Na+- channel scorpion toxins. PloS One. 2012;7 e30478. http://dx.doi.org/10.1371/journal.pone.0030478

Schwartz EF, Diego-García E, Rodríguez de la Vega RC, Possani LD. Transcriptome analysis of the venom gland of the Mexican scorpion Hadrurus gertschi (Arachnida: Scorpiones). BMC Genomics. 2007;8:119. http://dx.doi.org/10.1186/1471-2164-8-119

Chen T, Folan R, Kwok H, O’Kane EJ, Bjourson AJ, Shaw C. Isolation of scorpion (Androctonus amoreuxi) putative alpha neurotoxins and parallel cloning of their respective cDNAs from a single sample of venom. Regul Pept. 2003;115:115-21. http://dx.doi.org/10.1016/S0167-0115(03)00146-0

Chen T, Walker B, Zhou M, Shaw C. Molecular cloning of a novel putative potassium channel-blocking neurotoxin from the venom of the North African scorpion, Androctonus amoreuxi. Peptides. 2005;26:731-6. http://dx.doi.org/10.1016/j.peptides.2004.12.002

Jalali A, Bosmans F, Amininasab M, Clynen E, Cuypers E, Zaremirakabadi A, et al. OD1, the first toxin isolated from the venom of the scorpion Odontobuthus doriae active on voltage-gated Na+ channels. FEBS Letters. 2005;579:4181-6. http://dx.doi.org/10.1016/j.febslet.2005.06.052

Tytgat J, Chandy KG, García ML, Gutman GA, Martin-Eauclaire MF, van der Walt J, et al. A unified nomenclature for short-chain peptides isolated from scorpion venoms: Alpha-KTx molecular subfamilies. Trends Pharmacol Sci. 1999;20:444-7. http://dx.doi.org/10.1016/S0165-6147(99) 01398-X

Ortiz E, Gurrola GB, Schwartz EF, Possani LD. Scorpion venom components as potential candidates for drug development. Toxicon. 2015;93:125-35. http://dx.doi.org/10.1016/j.toxicon.2014.11.233

Zeng X, Corzo G, Hahin R. Scorpion venom peptideswithout disulfide bridges. IUBMB Life. 2005;57:13-21. http://dx.doi.org/10.1080/15216540500058899

Zeng XC, Li WX, Peng F, Zhu ZH. Cloning and characterization of a novel cDNA sequence encoding the precursor of a novel venom peptide (BmKbpp) related to a bradykininpotentiating peptide from Chinese scorpion Buthus martensii Karsch. IUBMB Life. 2000;49:207-10. http://dx.doi.org/10.1080/713803610

Almaaytah A, Albalas Q. Scorpion venom with no disulfide bridges: A review. Peptides. 2014;51:35-45. http://dx.doi.org/ 10.1016/j.peptides.2013.10.021

D’Suze G, Rosales A, Salazar V, Sevcik C. Apoptogenic peptides from Tityus discrepeans scorpion venom acting against the SKBR3 breast cancer cell line. Toxicon. 2010;56: 1497-505. http://dx.doi.org/10.1016/j.toxicon.2010.09.008

Guo X, Ma C, Du Q, Wei R, Wang L, Zhou M, et al. Two peptides, TsAP-1 and TsAP- 2, from the venom of the Brazilian yellow scorpion, Tityus serrulatus: Evaluation of their antimicrobial and anticancer activities. Biochimie. 2013;95:1784-94. http://dx.doi.org/10.1016/j.biochi.2013.06.003

Díaz A, Morier L, Rodríguez H, Caballero Y. Citotoxicidad del veneno del escorpión cubano Rhopalurus junceus y sus fracciones sobre líneas celulares tumorales humanas. LABIOFAM. 2010;1:12-8.

Rowe AH, Xiao Y, Scales J, Linse KD, Rowe MP, Cummins TR, et al. Isolation and characterization of CvIV4: A pain inducing α-scorpion toxin. PLoS One. 2011;6:e23520. http://dx.doi.org/10.1371/journal.pone.0023520

Das Gupta S, Debnath A, Saha A, Giri B, Tripathi G, Vedasiromoni J, et al. Indian black scorpion (Heterometrus bengalensis Koch) venom induced antiproliferative and apoptogenic activity against human leukemic cell lines U937 and K562. Leuk Res. 2007;31:817-25. http://dx.doi.org/10.1016/j.leukres.2006.06.004

Omran MA. In vitro anticancer effect of scorpion Leiurus quinquestriatus and Egyptian cobra venom on human breast and prostate cancer cell lines. J Med Sci. 2003;3:66-8. http://dx.doi.org/10.3923/jms.2003.66.86

Wang W, Ji Y. Scorpion venom induces glioma cell apoptosis in vitro and inhibits glioma tumor growth in vivo. J Neurooncol. 2005;73:1-7. http://dx.doi.org/10.1007/s11060-004-4205-6

Teruel R, García LF. Rare or poorly known scorpion from Colombia. I. Redescription of Tityus macrochirus Pocock, 1897. Euscorpius. 2008;63:1-11.

Oukkache N, Chgoury F, Lalaoui M, Cano A, Ghalim N. Comparison between two methods of scorpion venom milking in Morocco. J Venom Anim Toxins Incl Trop Dis. 2013;19:5. http://dx.doi.org/10.1186/1678-9199-19-5

Smith P, Krohn R, Hermanson G, Mallia AK, Gartner FH, Provenzano MD, et al. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985;150:76-85. http://dx.doi.org/10.1016/0003-2697(85)90442-7

Fernández E, Galván A. Métodos para la cuantificación de proteínas. Fecha de consulta: 12 de diciembre de 2015. Disponible en: http://www.uco.es/dptos/bioquimicabiol-mol/pdfs/27%20M%C3%89TODOS%20PARA%20LA%20CUANTIFICACI%C3%93N%20DE%20PROTE%C3%8DNAS.pdf.

Schagger H, von Jagow G. Tricine-sodium dodecyl sulfatepolyacrylamide

gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem. 1987;166:368-79. http://dx.doi.org/10.1016/0003-2697(87)90587-2

Pimenta A, Martin-Eauclaired MF, Rochat H, Figueiredo S, Kalapothakis E, Afonso LC, et al. Purification, aminoacid sequence and partial characterization of two toxins with anti-insect activity from the venom of the South American scorpion Tityus bahiensis (Buthidae). Toxicon. 200;39:1009-19. http://dx.doi.org/10.1016/S0041-0101(00)00240-3

Zargan J, Umar S, Sajad M, Naime M, Ali S, Haider A. Scorpion venom (Odontobuthus doriae) induces apoptosis by depolarization of mitochondria and reduces S-phase population in human breast cancer cells (MCF-7). Toxicol In Vitro. 2011;25:1748-56. http://dx.doi.org/10.1016/j.tiv.2011.09.002

ATCC. The essentials of life science research. Globally Delivered. Fecha de consulta: 27 de abril 27 de 2016. Disponible en: https://www.atcc.org/~/media/PDFs/QC_Strains.ashx.

Mosmann T. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J Immunol Methods. 1983;65:55-63. http://dx.doi.org/10.1016/0022-1759(83)90303-4

Borgues A, De Sousa L, Espinoza J, Santos R-G, Kalapothakis E, Valadares D, et al. Characterization of Tityus scorpion venoms using synaptosome binding assays and reactivity towards Venezuelan and Brazilian antivenoms. Toxicon. 2008;51:66-79. http://dx.doi.org/10.1016/j.toxicon.2007.08.006

Schwartz EF, Capes EM, Diego-García E, Zamudio F, Fuentes O, Possani LD, et al. Characterization of hadrucalcin, a peptide from Hadrurus gertschi scorpion venom with pharmacological activity on ryanodine receptors. Br J Pharmacol. 2009;157:392-403. http://dx.doi.org/10.1111/j.1476-5381.2009.00147.x

De Roodt AR. Veneno de escorpiones (alacranes) y envenenamiento. Acta Bioquím Clín Latinoam. 2015;49:55-71.

Kawachi T, Miyashita M, Nakagawa Y, Miyagawa H. Isolation and characterization of anti-insect β-toxin from venom the scorpion Isometrus maculatus. Biosci Biotechnol. 2013;77:205-507. http://dx.doi.org/10.1271/bbb.120697

Jäger H, Dreker T, Buck A, Giehl K, Gress T, Grissmer S. Blokage of intermediate-conductance Ca2+ activated K+ channels inhibit human pancreatic cancer cell growth in vitro. Mol Pharmacol. 2004;65:630-8. http://dx.doi.org/10.1124/mol.65.3.630

How to Cite
1.
Rincón-Cortés CA, Reyes-Montaño EA, Vega-Castro NA. Partial purification of peptides present in the Tityus macrochirus (Buthidae) scorpion venom and preliminary assessment of their cytotoxicity. biomedica [Internet]. 2017 Jun. 1 [cited 2024 May 13];37(2):238-49. Available from: https://revistabiomedica.org/index.php/biomedica/article/view/3265

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Published
2017-06-01
Section
Original articles

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