Abstract
The selective toxicity of depressant scorpion neurotoxins to insects is useful in studying the insect sodium channel gating, as well as being relevant to several other applications. In order to carry out structure/activity studies, the functional expression of such polypeptides is required. In the work reported here, the cDNA of a new peptide from the venom of the scorpion Buthotus saulcyi was cloned and sequenced. It codes for a 64 residues peptide (BsaulI) with 8 highly-conserved cysteines. This peptide shares high sequence similarity with depressant insect toxins of other scorpion species. Large amounts of insoluble BsaulI protein were expressed in Escherichia coli. Purification of this peptide was carried out under denaturing conditions. Renaturation was performed by pulsed dilution of the denatured BsaulI in the refolding buffer. Production of refolded Bsaul1, however, is approximately an order of magnitude higher than that obtained with similar scorpion depressant toxins. Intrinsic fluorescence, far-UV circular dichroism spectra and biological activity assays indicate that the peptide adopts a folded structure.
Keywords: Scorpion, depressant toxin, protein expression, refolding, Buthotus saulcyi, secondary structure
Protein & Peptide Letters
Title: Efficient In Vitro Refolding and Characterization of a New Peptide from the Scorpion Buthotus saulcyi Venom Produced in Escherichia coli
Volume: 13 Issue: 7
Author(s): Maryam Nikkhah, Hossein Naderi-Manesh, Mohamad N. Sarbolouki and Bijan Ranjbar
Affiliation:
Keywords: Scorpion, depressant toxin, protein expression, refolding, Buthotus saulcyi, secondary structure
Abstract: The selective toxicity of depressant scorpion neurotoxins to insects is useful in studying the insect sodium channel gating, as well as being relevant to several other applications. In order to carry out structure/activity studies, the functional expression of such polypeptides is required. In the work reported here, the cDNA of a new peptide from the venom of the scorpion Buthotus saulcyi was cloned and sequenced. It codes for a 64 residues peptide (BsaulI) with 8 highly-conserved cysteines. This peptide shares high sequence similarity with depressant insect toxins of other scorpion species. Large amounts of insoluble BsaulI protein were expressed in Escherichia coli. Purification of this peptide was carried out under denaturing conditions. Renaturation was performed by pulsed dilution of the denatured BsaulI in the refolding buffer. Production of refolded Bsaul1, however, is approximately an order of magnitude higher than that obtained with similar scorpion depressant toxins. Intrinsic fluorescence, far-UV circular dichroism spectra and biological activity assays indicate that the peptide adopts a folded structure.
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Cite this article as:
Nikkhah Maryam, Naderi-Manesh Hossein, Sarbolouki N. Mohamad and Ranjbar Bijan, Efficient In Vitro Refolding and Characterization of a New Peptide from the Scorpion Buthotus saulcyi Venom Produced in Escherichia coli, Protein & Peptide Letters 2006; 13 (7) . https://dx.doi.org/10.2174/092986606777790557
DOI https://dx.doi.org/10.2174/092986606777790557 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
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