Scorpion venom contains a group of two-domain peptides that function to block potassium channels or have cytolytic activities. These peptides, whose functions are poorly studied, are named beta-KTx or scorpine-like peptides. Ev37, the first identified gene in the Euscorpiidae family, which encoded a novel scorpine-like peptide, was cloned from the venom cDNA library of scorpion Euscorpiops validus. Sequence analysis showed that the mature Ev37 peptide contained 78 amino acid residues, which formed two structural domains: a putative alpha-helical N-terminus and a C-terminus with the cysteine-stabilized alpha/beta motif. The peptide rEv37 and two truncated peptides representing the individual structural domains (Ev37-N and Ev37-C) were expressed in Escherichia coli and purified for functional study. Unlike classic scorpine-like peptides, rEv37 and truncated peptides showed no cytolytic activity against bacteria or eukaryotic cells. Interestingly, rEv37 selectively inhibited Kv1.3 channel without effectively blocking Kv1.1 and Kv1.2 channels. Neither Ev37-N nor Ev37-C blocked Kv1.3 channel, suggesting that both the N-terminal and C-terminal domain of Ev37 are likely involved in the interaction with Kv1.3 channel. These results not only enrich our knowledge of scorpion toxins from scorpine-like subfamily but also provide a novel template with unique structure for designing new types of selective Kv1.3 blockers. (c) 2012 Elsevier Inc. All rights reserved.