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Soluble expression, purification and functional identification of a disulfide-rich conotoxin derived from Conus litteratus ☆

Paper ID Volume ID Publish Year Pages File Format Full-Text
25428 43573 2007 10 PDF Available
Title
Soluble expression, purification and functional identification of a disulfide-rich conotoxin derived from Conus litteratus ☆
Abstract

Conotoxins are a diverse array of small peptides mostly with multiple disulfide bridges. These peptides become an increasing significant source of neuro-pharmacological probes and drugs as a result of the high selectivity for ion channels and receptors. Usually, the analogue of natural conotoxins is produced by means of chemical synthesis. Here, we present a simple and fast strategy of producing disulfide-rich conotoxins via recombinant expression. By fused with thioredoxin and His tag, a novel O-superfamily conotoxin lt7a was successfully expressed in Escherichia coli and purified, resulting in a high yield of recombinant lt7a about 6 mg/l. The purity of target protein is up to 95% as identified by HPLC results. Whole cell patch-clamp recording revealed that the new conotoxin blocked voltage-sensitive sodium channels in rat dorsal root ganglion neurons, indicating it might be a novel μO-conotoxin.

Keywords
ACN, acetonitrile; BCA, bicinchoninic acid; DRG, dorsal root ganglion; HEPES, 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; PNK, polynucleotide kinase; TFA, trifluoroacetic acid; TRX, thioredoxin; VGSC, voltage-gated sodium channelCone snail; Conoto
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Soluble expression, purification and functional identification of a disulfide-rich conotoxin derived from Conus litteratus ☆
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 128, Issue 1, 30 January 2007, Pages 184–193
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us