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Keywords:

Amino Acid Sequence, Animals, Base Sequence, Cation Transport Proteins, Cells, Cultured, Cloning, Molecular, Electric Conductivity, Ether-A-Go-Go Potassium Channels, Models, Molecular, Molecular Sequence Data, Mutagenesis, Oocytes, Patch-Clamp Techniques, Phylogeny, Potassium Channel Blockers, Potassium Channels, Potassium Channels, Voltage-Gated, Protein Structure, Secondary, Scorpion Venoms, Xenopus laevis, Science & Technology, Life Sciences & Biomedicine, Biochemistry & Molecular Biology, Buthus martensi Karsch, gene cloning, phylogenetic tree, potassium channel, structure modelling, scorpion toxin, BUTHUS-MARTENSI KARSCH, POTASSIUM CHANNEL, GENOMIC ORGANIZATION, ANDROCTONUS-AUSTRALIS, CARDIAC-ARRHYTHMIA, BINDING-SITE, GENE, PEPTIDE, CHARYBDOTOXIN, RESIDUES, 03 Chemical Sciences, 06 Biological Sciences, 11 Medical and Health Sciences, 3101 Biochemistry and cell biology

Abstract:

A novel HERG channel blocker was isolated from the venom of the scorpion Buthus martensi Karsch, sequenced and characterized at the pharmacological level after chemical synthesis. According to the determined amino acid sequence, the cDNA and genomic genes were then cloned. The genomic gene consists of two exons interrupted by an intron of 65 bp at position -6 upstream from the mature toxin. The protein sequence of this toxin was completely identical with that of a known A-type K+ current blocker BmTx3, belonging to scorpion alpha-KTx subfamily 15. Thus BmTx3 is the first reported alpha-KTx peptide also showing HERG-blocking activity, like gamma-KTx peptides. Moreover, different from classical alpha-KTx peptides, such as charybdotoxin, BmTx3 cannot block Shaker -type K+ channels. Phylogenetic tree analysis reveals that this toxin takes an intermediate position between classical alpha-KTx and gamma-KTx toxins. From a structural point of view, we propose that two separate functional faces might exist on the BmTx3 molecule, responsible for the two different K+-current-blocking functions. Face A, composed of Arg18 and Lys19 in the alpha-helix side, might correspond to HERG blocking activity, whereas Face B, containing a putative functional dyad (Lys27 and Tyr36) in the beta-sheet side, might correspond to A-type blocking activity. A specific deletion mutant with the disrupted Face B, BmTx3-Y36P37del, loses the A-type current-blocking activity, but keeps a similar HERG-blocking activity, as seen with the wild-type toxin.