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^*

^*Korrespondierende/r Autor/-in für diese Arbeit

Institut für Experimentelle und Klinische Pharmakologie und Toxikologie

20 Zitate (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.

Originalsprache	Englisch
Zeitschrift	Protein Expression and Purification
Jahrgang	47
Ausgabenummer	2
Seiten (von - bis)	640-644
Seitenumfang	5
ISSN	1046-5928
DOIs	https://doi.org/10.1016/j.pep.2006.01.019
Publikationsstatus	Veröffentlicht - 01.06.2006

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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.

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Forschungsschwerpunkt: Gehirn, Hormone, Verhalten - Center for Brain, Behavior and Metabolism (CBBM)

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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.",

author = "Monika Bayrhuber and Roland Graf and Michael Ferber and Markus Zweckstetter and Julita Imperial and Garrett, \{James E.\} and Olivera, \{Baldomero M.\} and Heinrich Terlau and Stefan Becker",

year = "2006",

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doi = "10.1016/j.pep.2006.01.019",

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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

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SN - 1046-5928

VL - 47

SP - 640

EP - 644

JO - Protein Expression and Purification

JF - Protein Expression and Purification

IS - 2

ER -

Production of recombinant Conkunitzin-S1 in Escherichia coli

Abstract

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