Abstract
A gene, rBmK Cta, encoding a chlorotoxin-like peptide from the scorpion, Buthus martensii Karsch, was synthesized according to the sequence optimized for codon usage in Escherichia coli and was expressed in E.␣coli BL21 (DE3) using a pExSecI expression system in which the IgG-binding domain-ZZ of protein A is fused to the N-terminal of rBmK CTa. The fusion protein, ZZ-rBmK CTa, was expressed in soluble form (7.8 mg l−1) and was purified to give a single band on SDS-PAGE. The domain-ZZ of fusion protein ZZ-rBmK CTa was removed by cleavage of an Asn–Gly peptide bond with hydroxylamine. The rBmK CTa was separated from the IgG-binding moiety by a second passage through the IgG affinity column. Western blot analysis demonstrated that this protein was rBmK CTa. Acute toxicity assay in mice demonstrated that the rBmK CTa had an LD50 value of 4.3 mg kg−1.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
PE Bougis H Rochat LA Smith (1989) ArticleTitlePrecursors of Androctonus australis scorpion neurotoxins: structures of precursors, processing outcomes, and expression of a functional toxin II J. Biol. Chem. 264 19259–19265 Occurrence Handle2808423
C Brünen-Nieweler F Meyer K Heckmann (1994) ArticleTitleExpression of the pheromone 3-encoding gene of Euplotes octocarinatus using a novel bacterial secretion vector Gene 150 187–192 Occurrence Handle10.1016/0378-1119(94)90882-6 Occurrence Handle7959050
LF Carbonell MR Hodge MD Tomalski LK Miller (1998) ArticleTitleSynthesis of a gene coding for an insect-specific scorpion neurotoxin and attempts to express it using baculovirus vectors Gene 73 409–418 Occurrence Handle10.1016/0378-1119(88)90505-7
AR Dee M Belagaje K Ward E Chio MHT Lai (1990) ArticleTitleExpression and secretion of a functional scorpion insecticidal toxin in cultured mouse cells Bio/Technology 8 339–342 Occurrence Handle10.1038/ng0490-339 Occurrence Handle1366462
JA DeBin JE Maggio GR Strichartz (1993) ArticleTitlePurification and characterization of chlorotoxin, a chloride channel ligand from the venom of the scorpion Am. J. Physiol. 264 361–369
S Feng YM Xiong RH Zhu MH Ling CW Chi DC Wang (1999) ArticleTitleExpression and purification of the BmK M1 neurotoxin from the scorpion Buthus martensii Karsch Protein Expres. Purifi. 17 358–365 Occurrence Handle10.1006/prep.1999.1127
YJ Fu BF Chai W Wang H Zhi LT Yin AH Liang (2004) ArticleTitleExpression and purification of the BmK Mm2 neurotoxin from the scorpion Buthus martensii Karsch and its biological activity test Protein Expres. Purif. 38 45–50 Occurrence Handle10.1016/j.pep.2004.06.038
C Goudet CW Chi J Tytagt (2002) ArticleTitleAn overview of toxins and genes from the venom of the Asian scorpion Buthus martensii Karsch Toxicon 40 1239–1258 Occurrence Handle10.1016/S0041-0101(02)00142-3 Occurrence Handle12220709
YH Ji YJ Li JW Zhang BL Song Y Toshio M Tohru H Minoru Y Noboru (1999) ArticleTitleCovalent structures of BmK AS and BmK AS−1, two novel bioactive polypeptides purified from Chinese scorpion Buthus martrnsii Karsch Toxicon 37 519–536 Occurrence Handle10.1016/S0041-0101(98)00190-1 Occurrence Handle10080355
LY Jia HF Xie YH Ji (2000) ArticleTitleCharacterization of four distinct monoclonal antibodies specific to BmK AS-1, a novel scorpion bioactive polypeptide Toxicon 38 605–617 Occurrence Handle10.1016/S0041-0101(99)00175-0 Occurrence Handle10673154
C Maertens L Wei J Tytgat G Droogmans B Nilius (2000) ArticleTitleChlorotoxin does not inhibit volume-regulated, calcium-activated and cyclic AMP-activated chloride channels Br. J. Pharmacol. 129 791–801 Occurrence Handle10.1038/sj.bjp.0703102 Occurrence Handle10683204
MF Martin-Eauclaire M Sogaard C Ramos S Cestele PE Bougis B Svensson (1994) ArticleTitleProduction of active, insect-specific scorpion neurotoxin in yeast Eur. J. Biochem. 233 637–645 Occurrence Handle10.1111/j.1432-1033.1994.tb19036.x
J Meier RDG Theakston (1986) ArticleTitleApproximate LD50 determinations of snake venoms using eight to ten experimental animals Toxicon 24 395–401 Occurrence Handle10.1016/0041-0101(86)90199-6 Occurrence Handle3715904
T Moks L Abrahmsén E Holmgren M Bilich A Olsson M Uhlén G Pohl C Sterky H Hutberg S Josephson A Holmgern H Jǒrnvall B Nilsson (1987) ArticleTitleExpression of human insulin-like growth factor I in bacteria: use of optimized gene fusion vectors to facilitate protein purification Biochemistry 26 5239–5244 Occurrence Handle10.1021/bi00391a005 Occurrence Handle3676250
SZ Pang SM Oberhaus JL Rasmussen DC Knipple JF Bloomquist DH Dean KD Bowman JC Sanford (1992) ArticleTitleExpression of a gene encoding a scorpion insectotoxin peptide in yeast, bacteria and plants Gene 116 165–172 Occurrence Handle10.1016/0378-1119(92)90512-N Occurrence Handle1634114
JJ Wu D Li ZD Lan CW Chi (2000) ArticleTitleThe gene cloning and sequencing of Bm-12, a Chlorotoxin-like peptide from the scorpion Buthus martensii Karsch Toxicon 38 661–668 Occurrence Handle10.1016/S0041-0101(99)00181-6 Occurrence Handle10673158
YM Xiong MH Ling DC Wang CW Chi (1997) ArticleTitleThe cDNA and genomic DNA sequences of a mammalian neurotoxin from the scorpion Buthus martensii Karsch Toxicon 35 1025–1031 Occurrence Handle10.1016/S0041-0101(96)00224-3 Occurrence Handle9248001
XC Zeng WX Li SY Zhu F Peng ZH Zhu KL Wu FH Yang (2000) ArticleTitleCloning and characterization of a cDNA sequence encoding the precursor of a chlorotoxin-like peptide from the Chinese scorpion Buthus martensii Karsch Toxicon 38 1009–1014 Occurrence Handle10.1016/S0041-0101(99)00212-3 Occurrence Handle10708793
N Zilberberg D Gordon M Pelhate ME Adams TM Norris E Zlotkin M Gurevitz (1996) ArticleTitleFunctional expression and genetic alteration of an alpha scorpion neurotoxin Biochemistry 35 10215–10222 Occurrence Handle10.1021/bi9528309 Occurrence Handle8756487
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Fu, Yj., Yin, Lt., Wang, W. et al. Synthesis, Expression and Purification of a Type of Chlorotoxin-like Peptide from the Scorpion, Buthus martensii Karsch, and its Acute Toxicity Analysis. Biotechnol Lett 27, 1597–1603 (2005). https://doi.org/10.1007/s10529-005-2514-2
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s10529-005-2514-2