title: Efficient in vitro refolding and characterization of a new peptide from the scorpion Buthotus saulcyi venom produced in Escherichia coli.
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.
authors.affiliation: Institute of Biochemistry and Biophysics, University of Tehran, P.O. Box: 13145-1384, Tehran, Iran.
authors.forename: Maryam
authors.initials: M
authors.lastname: Nikkhah
authors.forename: Hossein
authors.initials: H
authors.lastname: Naderi-Manesh
authors.forename: Mohamad N
authors.initials: MN
authors.lastname: Sarbolouki
authors.forename: Bijan
authors.initials: B
authors.lastname: Ranjbar
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017510:Protein Folding; D012604:Scorpion Venoms / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D012605:Scorpions / Q000235:genetics / Q000378:metabolism*
chemical_list: D010455:Peptides; D012604:Scorpion Venoms