Production of recombinant Conkunitzin-S1 in Escherichia coli.
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.
Department of NMR based Structural Biology, Max-Planck-Institute for Biophysical Chemistry, Göttingen, Germany.
Monika
M
Bayrhuber
Roland
R
Graf
Michael
M
Ferber
Markus
M
Zweckstetter
Julita
J
Imperial
James E
JE
Garrett
Baldomero M
BM
Olivera
Heinrich
H
Terlau
Stefan
S
Becker
D000818:Animals; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D006863:Hydrogen-Ion Concentration; D002479:Inclusion Bodies / Q000378:metabolism; D047668:Inteins / Q000235:genetics; D008978:Mollusk Venoms / Q000096:biosynthesis* / Q000737:chemistry; D009865:Oocytes / Q000166:cytology / Q000378:metabolism; D018408:Patch-Clamp Techniques; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry; D051637:Shaker Superfamily of Potassium Channels / Q000037:antagonists & inhibitors; D014981:Xenopus
D008978:Mollusk Venoms; D011993:Recombinant Fusion Proteins; D051637:Shaker Superfamily of Potassium Channels; C504068:conkunitzin-S1, Conus striatus