Parabutoporin--an antibiotic peptide from scorpion venom--can both induce activation and inhibition of granulocyte cell functions.

Parabutoporin (PP) affects motility and NADPH oxidase activity in normal human polymorphonuclear neutrophils and in granulocytic HL-60 cells. These PP-induced interactions utilize a Rac activation pathway. PP induces chemotaxis of neutrophils and HL-60 cells via a pertussis toxin-sensitive way, thus using trimeric G-proteins. The enhanced chemotaxis is also apparent in undifferentiated HL-60 cells which lack functional formyl peptide receptors. On the other hand, PP strongly reduces the superoxide production by the NADPH oxidase complex after either PMA or fMLP activation of granulocytes. These combined results strongly suggest a direct activation of G-proteins and subsequent Rac activation as the basis for the observed effects. The unexpected inhibitory effect of PP, despite Rac activation, on superoxide production in granulocytes is explained by the direct interaction of membrane localized PP which prevents the formation of a functional NADPH oxidase complex.

Antimicrobial peptides from scorpion venom induce Ca(2+) signaling in HL-60 cells.

Parabutoporin (PP) and opistoporin 1 (OP1) are amphipathic alpha-helical antimicrobial peptides that were recently isolated from scorpion venom. In assays in which single granulocyte-like HL-60 cells as well as cells in suspension were used, both peptides were able to induce a reversible Ca(2+) release from intracellular stores and to increase Ca(2+) influx. Both effects could be clearly differentiated for OP1, inducing Ca(2+) release at lower concentrations. The Ca(2+) release was pertussis toxin-sensitive indicating the involvement of G-proteins. Ca(2+) release depended on the stage of differentiation of the cells with undifferentiated cells being the most sensitive. Desensitization occurred with OP1. No cross-desensitization occurred between OP1 and the bacterial chemoattractant fMLP indicating the involvement of different types of receptors. Ca(2+) release by OP1 was found not to be mediated via interaction with the formyl peptide receptor-like 1. Although some of the results might favor a receptor-like interaction, the receptor involved could not be identified.

Antibacterial and antifungal properties of alpha-helical, cationic peptides in the venom of scorpions from southern Africa.

Two novel pore-forming peptides have been isolated from the venom of the South-African scorpion Opistophtalmus carinatus. These peptides, designated opistoporin 1 and 2, differ by only one amino acid and belong to a group of alpha-helical, cationic peptides. For the first time, a comparison of the primary structures of alpha-helical pore-forming peptides from scorpion venom was undertaken. This analysis revealed that peptides in the range of 40-50 amino acids contain a typical scorpion conserved sequence S(x)3KxWxS(x)5L. An extensive study of biological activity of synthesized opistoporin 1 and parabutoporin, a pore-forming peptide previously isolated from the venom of the South-African scorpion Parabuthus schlechteri, was undertaken to investigate an eventual cell-selective effect of the peptides. Opistoporin 1 and parabutoporin were most active in inhibiting growth of Gram-negative bacteria (1.3-25 micro m), while melittin and mastoparan, two well-known cytolytic peptides, were more effective against Gram-positive bacteria in the same concentration range. In addition, the peptides showed synergistic activity with some antibiotics commonly used in therapy. Opistoporin 1 and parabutoporin had hemolytic activity intermediate between the least potent mastoparan and the highly lytic melittin. Furthermore, all peptides inhibited growth of fungi. Experiments with SYTOX green suggested that this effect is related to membrane permeabilization.