The venoms of Vespidae are rich in policationic peptides, generally involved in inflamatory proccesses, including cellular membrane lysis, mast cell degranulation (followed by histamine release), chemotaxis of neutrofils and T helper cells (causing recruitment of leukocytes to the site of stinging). The most abundant peptides found in wasp venoms are the mastoparans and chemotactic peptides. Among the diverse peptide components found in the venom of A. p. pallipes still there are some toxins not yet well characterized. In preliminary antibiotic assays the peptides Protonectin and Protonectin (1-6) presented a mutual interaction to each other at molecular level, potentiating the antimicrobial activity of Protonectin. The objectives of the present study is biochemical characterization of these peptides individually and also the supramolecular structure resulting from their interaction. The A. p. pallipes venom was extracted in MeCN 50% (v/v), purified by using HPLC under reverse phase and analyzed by mass spectrometry (ESI-MS and ESI-MS/MS). Both peptides were sequenced by Automated EDMAN Degradation Chemistry. Both peptides were manually synthesized on Solid Phase by using a Fmoc Chemistry. The peptides Protonectin and Protonectin (1-6) present molecular masses of 1210Da (I-L-G-T-I-L-G-L-L-K-G-L-NH2) and 628Da (I-L-G-T-I-L-NH2), respectively. In presence of TFE, Protonectin and Protonectin (1-6) tend to form 36.7% and 17.6% of a-helix, respectively; however, the mixture of both peptides (1:1) resulted in a supramolecular structure presenting 48.3% of a-helix. Protonectin alone presented chemotactic activity, hemolysis and poor mast cell degranulation, meanwhile Protonecitin (1-6) presented none of these activities. However, the equimolar mixture of both peptides resulted in a very potent activation of Protonectin activities, probably due to the formation of a supramolecular structure more organized than both peptides alone.

  Financial support: FAPESP/CNPq; iii/CNPq/MCT