Recombinant expression, purification, and characterization of scorpion toxin BmαTX14

Protein Expression and Purification
Hui DaiWenxin Li

Abstract

Long-chain and cysteine-rich scorpion toxins exhibit various pharmacological profiles for different voltage-gated sodium channel subtypes. However, the exploration of toxin structure-function relationships has progressed slowly due to the difficulty of obtaining synthetic or recombinant peptides. We now report that we have established an effective expression and purification approach for the novel scorpion toxin BmαTX14. BmαTX14 was over-expressed as inclusion bodies in Escherichia coli. The insoluble pellet was successfully transformed into active peptide by using a refolding procedure. One-step purification by reverse-phase HPLC was sufficient to generate chromatographically pure peptide. The yield of recombinant toxin reached 4mg from 1L LB medium. The pharmacological data further showed that BmαTX14 selectively inhibited the fast inactivation of mNa(v)1.4 (EC(50)=82.3±15.7nM) rather than that of rNa(v)1.2 (EC(50)>30μM), which indicates that BmαTX14 is a new α-like toxin. This work enables further structural, functional, and pharmacological studies of BmαTX14 and similar toxins.

References

Aug 5, 1998·Toxicon : Official Journal of the International Society on Toxinology·P SautièreD Gordon
Dec 22, 1999·Protein Expression and Purification·F ShaoD C Wang
Nov 22, 2000·Biochimie·S Cestèle, W A Catterall
Mar 26, 2003·FEBS Letters·Allan M TorresJamie I Vandenberg
May 11, 2004·The Journal of Biological Chemistry·Izhar KarbatMichael Gurevitz
Dec 1, 2004·Journal of Peptide Science : an Official Publication of the European Peptide Society·Sarrah M'BarekFrançois Sampieri
Jan 29, 2005·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Li-Hui LiuJan Tytgat
Feb 24, 2005·Biochimica Et Biophysica Acta·Christian LegrosMarie-France Martin-Eauclaire
Jul 9, 2005·Science·Stephen B LongRoderick Mackinnon
Nov 9, 2005·Toxicon : Official Journal of the International Society on Toxinology·Ricardo C Rodríguez de la Vega, Lourival D Possani
Jan 12, 2007·Toxicon : Official Journal of the International Society on Toxinology·Dalia GordonMichael Gurevitz
Jun 5, 2007·Biochimica Et Biophysica Acta·Georgina EstradaGerardo Corzo
Jun 27, 2007·Circulation·Argelia Medeiros-DomingoMichael J Ackerman
Jul 13, 2007·Biochemical and Biophysical Research Communications·Xin MaoWenxin Li
Aug 10, 2007·The Journal of Biological Chemistry·Lior CohenMichael Gurevitz
Jul 30, 2008·The Journal of General Physiology·Fabiana V CamposFrancisco Bezanilla
Feb 24, 2009·Toxicon : Official Journal of the International Society on Toxinology·Blanca I García-GómezLourival D Possani
Jun 11, 2009·The Journal of Biological Chemistry·Roy KahnMichael Gurevitz
Oct 13, 2009·Pain Medicine : the Official Journal of the American Academy of Pain Medicine·Sulayman D Dib-HajjStephen G Waxman
Jul 12, 2011·Nature·Jian PayandehWilliam A Catterall
Jul 19, 2011·Biochemical and Biophysical Research Communications·Bin GaoShunyi Zhu
Aug 31, 2011·Proceedings of the National Academy of Sciences of the United States of America·Jinti WangWilliam A Catterall

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Citations

Mar 1, 2014·Toxins·Zhijian CaoWenxin Li
Sep 10, 2014·Toxicon : Official Journal of the International Society on Toxinology·Lingna XuYingliang Wu
Nov 26, 2013·International Journal of Pharmaceutics·Hao HuKaichun Wu
Jun 3, 2018·Applied Microbiology and Biotechnology·Fernanda Gobbi AmorimEliane Candiani Arantes
Oct 30, 2016·Neuropharmacology·David M HousleyErnest A Jennings

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