Functional study of active residues scorpion insect toxin BmK IT from Buthus martensii Karsch

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• 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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• 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.