BmKb1

Innate immunity is the first line defense of multicellular organisms that rapidly operates to limit aggression upon exposure to pathogen microorganisms. Although the existence of some antibacterial peptides in scorpion venoms suggests that venom gland could be protected by these effector molecules, antibacterial activity of venom itself has not been assessed. In this study, we reported the antibacterial activity of the venom of Chinese scorpion Buthus martensii. Protease K digestion test indicated that it is venom peptide/protein components, as key players, which are involved in such antibacterial response. As the first step toward studying molecular mechanism of scorpion venom gland immunity, we established an infection model which supports inducible antibacterial response of scorpion venom gland. A known B. martensii antibacterial peptide gene BmKb1 was up-regulated at the transcriptional level after venom gland was challenged, suggesting its key defense role. This is further strengthened by the presence of several immune response elements in the BmKb1 promoter region. Our work thus provides the first evidence supporting the role of venom antibacterial peptides (ABPs) in controlling scorpion venom gland infection and lays a basis for characterizing related components involved in regulation of scorpion venom gland ABP gene expression.

At least 25 nondisulfide-bridged peptides (NDBPs) have been identified and characterized from scorpions. However, the genomic organization of the genes that encode these peptides have not been reported yet. BmKa1, BmKa2 and BmKb1 are three novel genes that code for NDBPs identified by our group from Mesobuthus martensii Karsch. Based on their cDNA sequences, the genomic DNA sequences encoding these peptides were obtained using the PCR method. Sequence analysis showed that three distinct genomic structural patterns are used to encode these three peptides. The BmKa1 gene is not interrupted by any introns. However, the BmKa2 gene is composed of two exons, interrupted by a 67 bp intron that is located in the DNA region encoding the mature peptide. Two genomic homologues of the BmKb1 cDNA sequence, named BmKb1' and BmKb2, respectively, were obtained. The BmKb1' gene contains one intron of 593 bp, inserted into the DNA region that encodes the signal peptide. Similarly, the BmKb2 gene also contains an intron that interrupts the exon that encodes the NDBP signal peptide. The amino acid sequences deduced for BmKb2 and BmKb1' differ only at one position. The data suggest that the genomic organizational pattern of NDBPs displays more divergence than that exhibited by the genes that encode disulfide-bridged peptides from scorpions.

The scorpion venom peptides with no disulfide bridge are rarely identified and poorly characterized so far. Here, we report the identification and characterization of four novel disulfide-bridge-free venom peptides (BmKa1, BmKa2, BmKb1 and BmKn2) from Buthus martensii Kasch. BmKa1 and BmKa2 are very acidic and hydrophilic, showing no any similarity to other proteins, whereas BmKb1 and BmKn2 both are basic, alpha-helical peptide with an amidated C-terminus, showing a little homology with other peptides. Functional tests with synthetic peptide showed that BmKn2 has strong antimicrobial activity against both Gram-positive and Gram-negative bacteria, whereas BmKb1 has weak activity in inhibiting the growth of these bacteria.