Design and construction of a chimeric peptide, MeICT/IMe-AGAP, from two anti-cancer toxins of Iranian Mesobuthus eupeus scorpion

Document Type : Original article

Authors

Department of Genetics, Shahrekord University, Rahbar Blvd, P.O Box 115 Shahrekord, Iran

Abstract

Scorpion venom contains various toxin peptides with pharmacological and biological properties. Scorpion toxins specifically interact with membrane ion channels which play key roles in progression of cancer. Therefore, scorpion toxins have received special attention for targeting cancer cells. Two new toxins MeICT and IMe-AGAP, isolated from Iranian yellow scorpion, Mesobuthus eupeus, interact specifically with chloride and sodium channels, respectively. Anti-cancer properties of MeICT and IMe-AGAP have been determined before, in addition they show 81 and 93% similarity with two well-known anti-cancer toxins, CTX and AGAP, respectively. The aim of this study was construction of a fusion peptide MeICT/IMe-AGAP to target different ion channels involved in cancer progression. Design and structure of the fusion peptide were investigated by bioinformatics studies. Two fragments encoding MeICT and IMe-AGAP were fused using overlapping primers by SOEing-PCR. MeICT/IMe-AGAP chimeric fragment was cloned into pET32Rh vector, expressed in Escherichia coli host and analyzed by SDS-PAGE. The in silico studies showed that chimeric peptide with GPSPG linker preserved the three-dimensional structure of both peptides and can be functional. Due to the high expression of chloride and sodium channels in various cancer cells, MeICT/IMe-AGAP fusion peptide can be used as an effective agent to target both channels in cancers, simultaneously.

Keywords

References

  1. Bordon KCF, Cologna CT, Fornari-Baldo EC, Pinheiro-Ju´ nior EL, Cerni FA, Amorim FG, Anjolette FAP, Cordeiro FA, Wiezel GA, Cardoso IA, Ferreira IG, Oliveira IS, Boldrini-Franc¸a J, Pucca MB, Baldo MA, Arantes EC. From animal poisons and venoms to medicines: achievements, challenges and perspectives in drug discovery. Front Pharmacol 2020;11:1132.
  2. Zhang XY, Zhang PY. Scorpion venoms in gastric cancer. Oncol Lett 2016;12:3683-3686.  
  3. Ortiz E, Possani LD. The unfulfilled promises of scorpion insectotoxins. J Venom Anim Toxins Incl Trop Dis 2015;21:16.
  4. Srairi-Abid N, Othman H, Aissaoui D, BenAissa R. Anti-tumoral effect of scorpion peptides: Emerging new cellular targets and signaling pathways. Cell Calcium 2019;80:160-174.
  5. Li M, Xiong ZG. Ion channels as targets for cancer therapy. Int J Physiol Pathophysiol Pharmacol 2011;3:156-166.
  6. Peretti M, Angelini M, Savalli N, Florio T, Yuspa SH, Mazzanti M. Chloride channels in cancer: Focus on chloride intracellular channel 1 and 4 (CLIC1 AND CLIC4) proteins in tumor development and as novel therapeutic targets. Biochim Biophys Acta 2015;1848: 2523-2531.
  7. Djamgoz M, Fraser SP, Brackenbury WJ. In vivo evidence for voltage-gated sodium channel expression in carcinomas and potentiation of metastasis. Cancers (Basel) 2019;11:1675.
  8. Ding J, Chua PJ, Bay BH, Gopalakrishnakone P. Scorpion venoms as a potential source of novel cancer therapeutic compounds. Exp Biol Med 2014;239:387-393.
  9. Ilkhanizade S, Ayat H, Ahadi A, Pirali K. Sequencing and comparative-bioinformatic analysis of chlorotoxin-like peptide from Iranian scorpion Mesobuthus eupeus. J Shahrekord Univ Med Sci 2011;13:27-36. [Persian]
  10. Dehghan Z, Ayat H, Ahadi AM. Bioinformatics analysis of analgesic-antitumor like peptide from Iranian scorpion Mesobuthus eupeus. J Shahrekord Univ Med Sci 2016;18:98-108. [Persian]
  11. Shahbazi Gandomkari M, Ayat H,  Ahadi AM. Recombinantly expressed MeICT, a new toxin from Mesobuthus eupeus scorpion, inhibits glioma cell proliferation and downregulates Annexin A2 and FOXM1 genes. Biotechnol Lett 2022;44:703-712. 
  12. Mohammadi Farsani F, Ayat H, Ahadi AM, Molecular Modeling and Docking Studies on the First Chlorotoxin-Like Peptide from Iranian Scorpion Mesobuthus eupeus (Meict) and SNP Variants of Matrix Methaloproteinase-2 (MMP-2). Iranian J Toxicol 2015;9:1368-1376.
  13. Cohen G, Burks SR, Frank JA. Chlorotoxin-A multimodal imaging platform for targeting glioma tumors. Toxins (Basel) 2018;10:496.
  14. Dehghan Z, Ayat H, Ahadi AM. Expression, purification and docking studies on IMe-AGAP, the first antitumor-analgesic like peptide from Iranian Scorpion Mesobuthus eupeus. Iran J Pharm Res 2020;19:206-216.
  15. Kampo S, Ahmmed B, Zhou T, Owusu L, Anabah TW, Doudou NR, Kuugbee ED, Cui Y, Lu Z, Yan Q, Wen QP. Scorpion venom analgesic peptide, BmK AGAP inhibits stemness, and epithelial-mesenchymal transition by down-regulating PTX3 in breast cancer. Front Oncol 2019;25;9:21.
  16. Gu Y, Liu SL, Ju WZ, Li CY, Cao P. Analgesic-antitumor peptide induces apoptosis and inhibits the proliferation of SW480 human colon cancer cells. Oncol Lett 2013;5:483-488.
  17. Bortner CD, Cidlowski J. Ion channels and apoptosis in cancer. Philos Trans R Soc Lond B Biol Sci 2014;369:20130104.
  18. Quintero-Hernández V, Jiménez-Vargas JM, Gurrola GB, Valdivia HH, Possani LD. Scorpion venom components that affect ion-channels function. Toxicon 2013;76:328-342.
  19. Djamgoz MBA, Fraser SP, Brackenbury WJ. In vivo evidence for voltage-gated sodium channel expression in carcinomas and potentiation of metastasis. Cancers (Basel) 2019; 11:1675.
  20. Chandy KG, Norton RS. Peptide blockers of Kv1.3 channels in T cells as therapeutics for autoimmune disease. Curr Opin Chem Biol 2017;38:97-107.
  21. Quintero-Fabián S, Arreola R, Becerril-Villanueva E, Torres-Romero JC, Arana-Argáez V, Lara-Riegos J, Ramírez-Camacho MA,  Alvarez-Sánchez ME. Role of matrix metallo-proteinasees in angiogenesis and cancer. Front Oncol 2019;9:1370.
  22. Song Z, Wang J, Su Q, Luan M, Chen X, Xu X. The role of MMP-2 and MMP-9 in the metastasis and development of hypopharyngeal carcinoma. Braz J Otorhinolaryngol 2021; 87:521-528.
  23. Srinivasa Rao K, Srinivas K, Sujini GN, Sunand Kumar GN. Protein-protein interaction detection: Methods and analysis. Int J Proteomics 2014;2014:147648. 
  24. Reddy Chichili VP, Kumar V, Sivaraman J. Linkers in the structural biology of protein-protein interactions. Protein Sci 2013;22:153-167.
  25. Dastpeyman M, Giacomin P, Wilson D, Nolan MJ. A C-terminal fragment of chlorotoxin retains bioactivity and inhibits cell migration. Pharmacol 2019;10:250.
  26. Zhang R, Cui Y, Zhang X, Yang Z, Zhao Y, Song YB, Wu C, Zhang J. Soluble expression, purification and the role of C-terminal glycine residues in scorpion toxin BmK AGP-SYPU2. BMB Rep 2010;43:801-806.
  27. Costa S, Almeida A, Castro A, Domingues L. Fusion tags for protein solubility, purification, and immunogenicity in Escherichia coli: the novel Fh8 system. Front Microbiol 2014;5:63.
  28. De Marco A. Strategies for successful recombinant expression of disulfide bond-dependent proteins in Escherichia coli. Microb Cell fact 2009;8:1-18.

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