Four disulfide-bridged scorpion beta neurotoxin CssII: Heterologous expression and proper folding in vitro
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Four disulfide-bridged scorpion beta neurotoxin CssII: heterologous expression and proper folding in vitro.
The gene of the four disulfide-bridged Centruroides suffusus suffusus toxin II was cloned into the expression vector pQE30 containing a 6His-tag and a FXa proteolytic cleavage region. This recombinant vector was transfected into Escherichia coli BL21 cells and expressed under induction with isopropyl thiogalactoside (IPTG). The level of expression was 24.6 mg/l of culture medium, and the His ta...
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Maurotoxin (MTX) is a 34-mer scorpion toxin cross-linked by four disulphide bridges that acts on various K(+) channel subtypes. MTX adopts a disulphide bridge organization of the type C1-C5, C2-C6, C3-C4 and C7-C8, and folds according to the common alpha/beta scaffold reported for other known scorpion toxins. Here we have investigated the process and kinetics of the in vitro oxidation/folding o...
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Scorpion venom is the richest source of peptide toxins with high levels of specific interactions with different ion-channel membrane proteins. The present study involved the amplification and sequencing of a 310-bp cDNA fragment encoding a beta-like neurotoxin active on sodium ion-channel from the venom glands of scorpion Androctonus crassicauda belonging to the Buthidae family using r...
متن کاملChaperone-mediated native folding of a β-scorpion toxin in the periplasm of Escherichia coli☆
BACKGROUND Animal neurotoxin peptides are valuable probes for investigating ion channel structure/function relationships and represent lead compounds for novel therapeutics and insecticides. However, misfolding and aggregation are common outcomes when toxins containing multiple disulfides are expressed in bacteria. METHODS The β-scorpion peptide toxin Bj-xtrIT from Hottentotta judaica and fou...
متن کاملThe disulfide bonds in antibody variable domains: effects on stability, folding in vitro, and functional expression in Escherichia coli.
The formation of the disulfide bonds in the variable domains VH and VL of the antibody McPC603 was found to be essential for the stability of all antigen binding fragments investigated. Exposure of the Fv fragment to reducing conditions in vitro resulted in irreversible denaturation of both VH and VL. In vitro refolding of the reduced Fv fragment was only possible when the disulfide bonds were ...
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ژورنال
عنوان ژورنال: Biochimica et Biophysica Acta (BBA) - General Subjects
سال: 2007
ISSN: 0304-4165
DOI: 10.1016/j.bbagen.2007.04.006