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Influence of a NH 2 ‐terminal extension on the activity of KTX2, a K + channel blocker purified from Androctonus australis scorpion venom
Author(s) -
Legros Christian,
Feyfant Eric,
Sampieri François,
Rochat Hervé,
Bougis Pierre E,
Martin-Eauclaire Marie-France
Publication year - 1997
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(97)01177-0
Subject(s) - venom , periplasmic space , scorpion toxin , escherichia coli , complementary dna , toxin , amino acid , biochemistry , scorpion venoms , fusion protein , biology , recombinant dna , chemistry , microbiology and biotechnology , stereochemistry , scorpion , gene
A cDNA encoding a short polypeptide blocker of K + channels, kaliotoxin 2 (KTX2), from the venom of the North African scorpion Androctonus australis was expressed in the periplasmic space of Escherichia coli . KTX2 was produced as a fusion protein with the maltose binding protein followed by the recognition site for factor Xa or enterokinase preceding the first amino acid residue of the toxin. The fully refolded recombinant KTX2 (rKTX2) was obtained (0.15–0.30 mg/l of culture) and was indistinguishable from the native toxin according to chemical and biological criteria. An N‐extended analogue of KTX2 exhibiting three additional residues was also expressed. This analogue had 1000‐fold less affinity for the 125 I‐kaliotoxin binding site on rat brain synaptosomes than KTX2. Conformational models of KTX2 and its mutant were designed by amino acid replacement using the structure of agitoxin 2 from Leiurus quinquestriatus as template, to try to understand the decrease in affinity for the receptor.