Chaperone-mediated native folding of a β-scorpion toxin in the periplasm of Escherichia coli.

ABSTRACT:

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 four chaperone enzymes (DsbA, DsbC, SurA and FkpA) were co-secreted into the oxidizing environment of the Escherichia coli periplasm. Expressed Bj-xtrIT was purified and analyzed by HPLC and FPLC chromatography. Its thermostability was assessed using synchrotron radiation circular dichroism spectroscopy and its crystal structure was determined.

Results

Western blot analysis showed th

SUBMITTER: O'Reilly AO 

PROVIDER: S-EPMC3898981 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature