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Published on: January 19, 2011
Expression of a functional recombinant Phoneutria nigriventer toxin active on K+ channels
A M D Carneiro1, C Kushmerick, J Koenen
1Departamento de Bioquímica-Imunologia, ICB, Universidade Federal de Minas Gerais, Av Antonio Carlos, 6627, Belo Horizonte, 31270-901, Minas Gerais, Brazil.
Abstract:
PnTx3-1 is a peptide isolated from the venom of the spider Phoneutria nigriventer that specifically inhibits A-type K(+) currents (I(A)) in GH(3) cells. Here we used a bacterial expression system to produce an NH(2)-extended mutant of PnTx3-1 (ISEF-PnTx3-1) and tested whether the toxin is functional. The recombinant toxin was purified from bacterial extracts by a combination of affinity and ion-exchange chromatography. The recombinant toxin blocked A-type K(+) currents in GH(3) cells in a fashion similar to that observed with the wild-type toxin purified from the spider venom. These results suggest that recombinant cDNA methods provide a novel source for the production of functional Phoneutria toxins. The recombinant ISEF-PnTx3-1 should be useful for further understanding of the role of A-type K(+) currents in biological processes.
