Production of recombinant Conkunitzin-S1 in Escherichia coli

被引：21

作者：

Bayrhuber, Monika

Graf, Roland

Ferber, Michael

Zweckstetter, Markus

Imperial, Julita

Garrett, James E.

Olivera, Baldomero M.

Terlau, Heinrich

Becker, Stefan

机构：

[1] Max Planck Inst Biophys Chem, Dept NMR Based Struct Biol, D-37077 Gottingen, Germany

[2] Univ Utah, Dept Biol, Salt Lake City, UT 84112 USA

[3] Cognetix Inc, Salt Lake City, UT 84108 USA

[4] Max Planck Inst Expt Med, Mol & Cellular Neuropharmacol Grp, D-37075 Gottingen, Germany

[5] Univ Clin Schleswig Holstein, Dept Expt & Clin Pharmacol & Toxicol, D-23538 Lubeck, Germany

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2006年 / 47卷 / 02期

关键词：

Conkunitzin-S1; Kunitz domain fold; cone snail; potassium channel;

D O I：

10.1016/j.pep.2006.01.019

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Conkunitzin-S1 from the cone snail Conus striatus is the first member of a new neurotoxin family with a canonical Kunitz domain fold. Conk-SI is 60 amino acids long and lacks one of the three conserved disulfide bonds typically found in Kunitz domain modules. It binds specifically to voltage activated potassium channels of the Shaker family. The peptide was expressed in insoluble form in fusion with an N-terminal intein. Refolding in the presence of glutathione followed by pH shift-induced cleavage of the fusion protein resulted in a functional toxin as demonstrated by voltage-clamp measurements. (c) 2006 Elsevier Inc. All rights reserved.

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页码：640 / 644

页数：5

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