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Functional study of active residues scorpion insect toxin BmK IT from Buthus martensii Karsch
- 作者:Yuejun Fu (1)
Renjia Yang (1) Wujian Chen (2) Zhiyi Wu (2) Aihua Liang (1) Fengyun Hu (3)
- 关键词:active residues ; Buthus martensii Karsch insect toxin (BmK IT) ; rat glioma C6 cell ; Spodopter frugiperda 9 cell ; cytotoxicity
- 刊名:Biotechnology and Bioprocess Engineering
- 出版年:2014
- 出版时间:March 2014
- 年:2014
- 卷:19
- 期:2
- 页码:320-326
- 全文大小:
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2. Liang, A. H., X. L. Li, and Z. G. Su (1999) Cloning and sequencing of an excitatory insect-selective neurotoxin / BmK IT cDNA from / Buthus martensii Karsch. / High Tech. Lett. 9: 12-5. 3. Hao, C. J., C. G. Xu, W. Wang, B. F. Chai, and A. H. Liang (2005) Expression of an insect excitatory toxin, / BmK IT, from the scorpion, / Buthus martensii Karsch, and its biological activity. / Biotechnol. Lett. 27: 1929-934. CrossRef 4. Fu, Y. J., R. J. Yang, A. H. Liang, C. G. Xu, and C. H. Hu (2011) Recombinant scorpion insect excitatory toxin / BmK IT accelerates the growth of insect Spodoptera frugiperda 9 cells. / Mol. Cell Biochem. 351: 93-8. CrossRef 5. Oren, D. A., O. Froy, E. Amit, N. Kleinberger-Doron, Gurevitz, and M. B. Shaanan (1998) An excitatory scorpion toxin with a distinctive feature: An additional alpha helix at the C terminus and its implications for interaction with insect sodium channels. / Structure 6: 1095-103. CrossRef 6. Li, C., R. J. Guan, Y. Xiang, Y. Zhang, and D. C. Wang (2005) Structure of an excitatory insect-specific toxin with an analgesic effect on mammals from the scorpion / Buthus martensii Karsch. / Acta Crystallogr. D. 61: 14-1. CrossRef 7. Tong, X., J. Yao, F. He, X. Chen, X. Zheng, C. Xie, G. Wu, N. Zhang, J. Ding, and H. Wu (2006) NMR solution structure of / BmK- / βIT, an excitatory scorpion beta-toxin without a ‘hot spot-at the relevant position. / Biochem. Biophys. Res. Co. 349: 890-99. CrossRef 8. Cohen, L., I. Karbat, N. Gilles, O. Froy, G. Corzo, R. Angelovici, D. Gordon, and M. Gurevitz (2004) Dissection of the functional surface of an anti-insect excitatory toxin illuminates a putative “hot spot-common to all scorpion beta-toxins affecting Na+ channels. / J. Biol. Chem. 279: 8206-211. CrossRef 9. Pedraza Escalona, M. and L. D. Possani (2013) Scorpion betatoxins and voltage-gated sodium channels: Interactions and effects. / Front Biosci. 18: 572-87. CrossRef 10. Guerrero-Vargas, J. A., C. B. Mour?o, V, Quintero-Hernández, L. D. Possani, and E. F. Schwartz (2012) Identification and phylogenetic analysis of Tityus pachyurus and Tityus obscurus novel putative Na+-channel scorpion toxins. / PLoS One 7: e30478. CrossRef 11. Gu, J. B., Y. Q. Dong, H. J. Peng, and X. G. Chen (2010) A recombinant AeDNA containing the insect-specific toxin, BmK IT1, displayed an increasing pathogenicity on Aedes albopictus. / Am. J. Trop. Med. Hyg. 83: 614-23. CrossRef 12. Xu, C. G., X. J. Fan, Y. J. Fu, and A. H. Liang (2008) Effect of location of the His-tag on the production of soluble and functional / Buthus martensii Karsch insect toxin. / Protein Expr. Purif. 59: 103-09. CrossRef 13. Ji, S. J., F. Liu, E. Q. Li, and Y. X. Zhu (2002) Recombinant scorpion insectotoxin AaIT kills specifically insect cells but not human cells. / Cell Res. 12: 143-50. CrossRef 14. Fu, Y. J., R. Huang, Y. L. Zheng, Z. Y. Zhang, and A. H. Liang (2011) Glioma-derived mutations in isocitrate dehydrogenase 2 beneficial to traditional chemotherapy. / Biochem. Bioph. Res. Co. 410: 218-23. CrossRef 15. Fu, Y. J., B. F. Chai, W. Wang, H. Zhi, L. T. Yin, and A. H. Liang (2004) Expression and purification of the BmK Mm2 neurotoxin from the scorpion / Buthus martensii Karsch and its biological activity test. / Protein Expres. Purif. 38: 45-0. CrossRef 16. Grobben, B., P. P. De Deyn, and H. Slegers (2002) Rat C6 glioma as experimental model system for the study of glioblastoma growth and invasion. / Cell Tissue Res. 310: 257-70. CrossRef 17. Billen, B., F. Bosmans, and J. Tytgat (2008) Animal peptides targeting voltage-activated sodium channels. / Curr. Pharm. Des. 14: 2492-502. CrossRef
- 作者单位:Yuejun Fu (1)
Renjia Yang (1) Wujian Chen (2) Zhiyi Wu (2) Aihua Liang (1) Fengyun Hu (3)
1. Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan, 030-006, China 2. Entry-Exit Inspection and Quarantine Bureau, Hangzhou, 310-012, China 3. Department of Neurology, Shanxi Provincial People’s Hospital, Taiyuan, 030-012, China
- ISSN:1976-3816
文摘
Chinese scorpion Buthus martensii Karsch (BmK) venom is a rich source of neurotoxins which bind to various ion channels with high affinity and specificity and thus widely used as compounds to modulate channel gating. An excitatory insect toxin, BmK IT, is not conserved with a glutamate residue at the preceding position of the third Cys residue, and is a toxin with a non-glutamate residue at the relevant position in the excitatory scorpion β-toxin subfamily. In this study, the mutants of recombinant BmK IT (BmK IT (I25E), BmK IT (E15G), BmK IT C-terminal (TKSYCDVQIN) truncated) were achieved by site-directed mutagenesis. Biological activity of BmK IT and its mutants confirmed these residues or peptides played key roles in BmK IT. BmK IT (I25E) could increase the sensitivity of BmK IT, but BmK IT(E15G) could decrease the sensitivity of BmK IT on Sf9 cells. BmK IT truncated C-terminal hydrophobic amino acids could cross the species boundaries and was effective on mammalian C6 cells. To date, several excitatory insect toxins have been isolated and identified from the venom of Buthus martensii Karsch. However, no functional data are available and therefore its classification in the family of excitatory insect toxins remains putative and is just based on its high similarity with the other toxins of this family. These results verified I25, E15 and C-terminal (TKSYCDVQIN) in BmK IT played key roles in the interaction of the BmK IT and its receptor- sodium channels on the surface of insect cells and laid a foundation for further structural and functional analysis of BmK IT.
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