Production of recombinant Conkunitzin-S1 in Escherichia coli

Monika Bayrhuber; Roland Graf; Michael Ferber; Markus Zweckstetter; Julita Imperial; James E. Garrett; Baldomero M. Olivera; Heinrich Terlau; Stefan Becker

doi:10.1016/j.pep.2006.01.019

Production of recombinant Conkunitzin-S1 in Escherichia coli

Monika Bayrhuber, Roland Graf, Michael Ferber, Markus Zweckstetter, Julita Imperial, James E. Garrett, Baldomero M. Olivera, Heinrich Terlau, Stefan Becker^*

^*Corresponding author for this work

Institute of Experimental and Clinical Pharmacology and Toxicology

20 Citations (Scopus)

Abstract

Conkunitzin-S1 from the cone snail Conus striatus is the first member of a new neurotoxin family with a canonical Kunitz domain fold. Conk-S1 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.

Original language	English
Journal	Protein Expression and Purification
Volume	47
Issue number	2
Pages (from-to)	640-644
Number of pages	5
ISSN	1046-5928
DOIs	https://doi.org/10.1016/j.pep.2006.01.019
Publication status	Published - 01.06.2006

Funding

We thank Petra Widawka, Kamila Budzyn, and Karin Giller for expert technical help and Christian Griesinger for useful discussions. M.Z. is supported by a DFG Emmy Noether Grant (Z.W. 71/1–5). J.I. and B.M.O. were supported by NIH Grant GM48677. This work was supported by the Max Planck Society.

Research Areas and Centers

Academic Focus: Center for Brain, Behavior and Metabolism (CBBM)

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.pep.2006.01.019

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title = "Production of recombinant Conkunitzin-S1 in Escherichia coli",

abstract = "Conkunitzin-S1 from the cone snail Conus striatus is the first member of a new neurotoxin family with a canonical Kunitz domain fold. Conk-S1 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.",

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AU - Bayrhuber, Monika

AU - Graf, Roland

AU - Ferber, Michael

AU - Zweckstetter, Markus

AU - Imperial, Julita

AU - Garrett, James E.

AU - Olivera, Baldomero M.

AU - Terlau, Heinrich

AU - Becker, Stefan

PY - 2006/6/1

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N2 - Conkunitzin-S1 from the cone snail Conus striatus is the first member of a new neurotoxin family with a canonical Kunitz domain fold. Conk-S1 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.

AB - Conkunitzin-S1 from the cone snail Conus striatus is the first member of a new neurotoxin family with a canonical Kunitz domain fold. Conk-S1 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.

UR - https://www.scopus.com/pages/publications/33646558307

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DO - 10.1016/j.pep.2006.01.019

M3 - Journal articles

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SN - 1046-5928

VL - 47

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EP - 644

JO - Protein Expression and Purification

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ER -

Production of recombinant Conkunitzin-S1 in Escherichia coli

Abstract

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Research Areas and Centers

UN SDGs

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