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Purification, characterization and cDNA cloning of an analgesic peptide from the chinese scorpion Buthus martensii Karsch (BmK AGP-SYPU2)

  • Genomics. Transcriptomics
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Abstract

In this study an analgesic peptide was purified through five continuous chromatographic steps. The mouse twisting model test was used to identify the target peptides in every separation step. The purified BmK AGP-SYPU2 was further qualified by Reverse Phase-High Performance Liquid Chromatography and High Performance Capillary Electrophoresis. The molecular weight, isoelectric point, and N-terminal sequence of the purified peptide were determined. Based on the N-terminal sequence, the cDNA was cloned by rapid amplification of the cDNA ends from the cDNA pool of scorpion glands. Sequence determination showed that the mature BmK AGP-SYPU2 peptide is composed of 66 amino acid residues, and BmK AGP-SYPU2 is identical to BmK α2 (GenBank Acc. No. AF288608) and BmK αTX11 (GenBank Acc. No. AF155364). We report herein a purification procedure that yields substantial amounts of natural BmK AGP-SYPU2 with high analgesic activity.

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Abbreviations

BmK:

Buthus martensii Karsch

IEC:

Ion exchange chromatography

HIC:

Hydrophobic interaction chromatography

GF:

Gel filtration chromatography

IEF:

Isoelectric focusing

HPCE:

High Performance Capillary Electrophoresis

References

  1. Liu Y.F., Ma R.L. Wang S.L., et al. 2003. Expression of an antitumor-analgesic peptide from the venom of Chinese scorpion Buthus martensii Karsch. in Escherichia coli. Protein Expr. Purif. 27, 253–258.

    Article  PubMed  Google Scholar 

  2. Li F., Lu S.N., Pan L.S. 1997. The experimental evaluation of scorpion toxins physical dependence. Chinese J. Pharmacol. Toxin. 11, 154–158.

    Google Scholar 

  3. Wang C.Y., Tan Z.Y., Chen B., et al. 2000. Antihyperalgesia effect of BmK IT2, a depressant insect-selective scorpion toxin in rat by peripheral administration. Brain Res. Bull. 53, 335–338.

    Article  PubMed  CAS  Google Scholar 

  4. Guan R.J., Liu X.Q., Liu B., et al. 2000. Crystallization and preliminary X-ray analyses of insect neurotoxins with analgesic effect from the scorpion Buthus martensii Karsch. Acta Crystallogr. D: Biol. Crystallogr. 56, 1012–1014.

    Article  CAS  Google Scholar 

  5. Guan R.J., Wang C.G., Wang M., et al. 2001. A depressant insect toxin with a novel analgesic effect from scorpion Buthus martensii Karsch. Biochim. Biophys. Acta. 1549, 9–18.

    Article  PubMed  CAS  Google Scholar 

  6. Lan Z.D., Dai L., Zhou X.L. 1999. Gene clone and sequencing of BmK AS and BmK AS-1, two novel neurotoxins from the scorpion Buthus martensi Karsch. Toxicon. 37, 815–823.

    Article  PubMed  CAS  Google Scholar 

  7. Guan R.J., Wang M., Wang D., et al. 2001. A new insect neurotoxin AngP1 with analgesic effect from the scorpion Buthus martensii Karsch: Purification and characterization. J. Pep. Res. 58, 27–35.

    Article  CAS  Google Scholar 

  8. Cao Z.Y., Mi Z.M., Cheng G.F., et al. 2004. Purification and characterization of a new peptide with analgesic effect from the scorpion Buthus martensi Karch. J. Pep. Res. 64, 33–41.

    Article  CAS  Google Scholar 

  9. Wang Y., Wang L., Cui Y., et al. 2010. Purification, characterization and functional expression of a new peptide with an analgesic effect from Chinese scorpion Buthus martensii Karsch (BmK AGP-SYPU1). Biomed. Chromatogr. 25, 801–807.

    Article  PubMed  Google Scholar 

  10. Fennessy M.R., Lee J.R. 1975. Methods in Narcotics Research. N.Y.: Marcel Dekker, pp. 76–79.

    Google Scholar 

  11. Bougis P.E., Rochat H., Smith L.A. 2006. Precursors of Androctonus australis Scorpion Neurotoxins. J. Biol. Chem. 264, 19259–19265.

    Google Scholar 

  12. Sambrook J., Fritsch E.E., Maniatis T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor, NY: Cold Spring Harbor Lab. Press.

    Google Scholar 

  13. Tan P.T., Veeramani A., Srinivasan K.N. 2006. SCORPION2: A database for structure-function analysis of scorpion toxins. Toxicon. 44, 758–764.

    Google Scholar 

  14. Shao J.H., Kang N., Liu Y.F., et al. 2007. Purification and characterization of an analgesic peptide from Buthus martensii Karsch. Biomed. Chromatogr. 21, 1266–1271.

    Article  PubMed  CAS  Google Scholar 

  15. Martin E., Couraud M.F. 1995. Scorpion neurotoxins: Effects and mechanisms. In: Handbook of Neurotoxicology. N.Y.: Marcel Dekker, pp. 683–716.

    Google Scholar 

  16. Wullschleger B., Nentwig W., Kuhn-Nentwig L. 2005. Spider venom: Enhancement of venom efficacy mediated by different synergistic strategies in Cupiennius salei. J. Exp. Biol. 208, 2115–2121.

    Article  PubMed  CAS  Google Scholar 

  17. Ye J.G., Chen J., Zuo X.P., et al. 2001. Cloning and characterization of cDNA sequences encoding two novel alpha-like-toxin precursors from the Chinese scorpion Buthus martensii Karsch. Toxicon. 39, 1191–1194.

    Article  PubMed  CAS  Google Scholar 

  18. Xu X.L., Cao Z.J., Sheng J.Q., et al. 2005. Genomic sequence analysis and organization of BmKalphaTx11 and BmKalphaTx15 from Buthus martensii Karsch: Molecular evolution of alpha-toxin genes. J. Biochem. Mol. Biol. 38, 386–390.

    Article  PubMed  CAS  Google Scholar 

  19. Brose W.G., Gutlove D.P. 1997. Use of intrathecal SNX-III, a novel, N-type, voltage-sensitive, calcium channel blocker in the management of intractable brachinal plexus arulsion pain. Clin. J. Pain. 13, 256–259.

    Article  PubMed  CAS  Google Scholar 

  20. Liu C.M., Pei G.Q. 1989. The analgesic effect research of scorpion venom from Buthus martensii Karsch. J. Shenyang Coll. Pharm. 6, 176–179.

    Google Scholar 

  21. Wang Y.K., Han X.F. 1996. The analgesic effect and its mechanism research of SAP in rat spinal medulla. J. Henan Med. University. 31, 5–8.

    Google Scholar 

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Authors and Affiliations

  1. School of Life Science and Bio-pharmaceutics, Shenyang Pharmaceutical University, 103 Wenhua Road, Shenyang, 110016, China

    R. Zhang, Z. Yang, Y. F. Liu, Y. Cui & J. H. Zhang

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  1. R. Zhang
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  2. Z. Yang
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  3. Y. F. Liu
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  4. Y. Cui
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  5. J. H. Zhang
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Correspondence to J. H. Zhang.

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Zhang, R., Yang, Z., Liu, Y.F. et al. Purification, characterization and cDNA cloning of an analgesic peptide from the chinese scorpion Buthus martensii Karsch (BmK AGP-SYPU2). Mol Biol 45, 879–885 (2011). https://doi.org/10.1134/S0026893311060203

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