On the other hand, CgNa is a toxin isolated from Condylactis gigantea species and presents a dense negative charge around residues 35–37 (see Table 1). In spite of the presence of such a negative charge its potency is in a similar range such as BgII when tested in dorsal root ganglia (DRG) neurons [28], [29] and [32]. Also, the determination of CgNa three-dimensional structure by NMR exhibits a large negative patch exposed, and

a minor distribution of hydrophobic residues that are important for activity in ApB and ATX-II [29]. As pointed by the authors, this may explain that at least for CgNa the presence of positively charged amino acids and a hydrophobic Z-VAD-FMK patch may not be of utmost importance for its binding on sodium channels, but might contribute to a smaller potency compared

to ATX-II and selleck inhibitor ApB, for instance. Observing the modeled structures shown in Fig. 5, we clearly see that for δ-AITX-Bcg1a peptide an overall charge distribution similar to CgNa may occur. In its primary sequence, we observe a negatively charged amino acid (D37) that is positioned in a correspondent region of D36 and E37 in CgNa, and their contact surface on sodium channels may also be similar. Comparing among the charged molecular surfaces of the three toxins, δ-AITX-Bcg1a has a more intense negatively charged surface when compared to CGTX-II. As for δ-AITX-Bcg1b, the occurrence of an Asp at position 16 may disrupt this possible surface of contact, by increasing the extent of the negative patch and make δ-AITX-Bcg1b much less prone to affect VGSC,

as shown in Fig. 1. This is especially interesting as we consider the only N16D substitution observed between δ-AITX-Bcg1a and δ-AITX-Bcg1b, but the molecular surface of the latter shows that the occurrence of Asp16 drastically increases the negatively charged surface among all three peptides. Consequently, we may suggest that the large negative surface observed in δ-AITX-Bcg1b may be responsible for its mild effects among the eltoprazine assayed Navs. In summary, our results contribute to a better understanding of the selectivity of some sea anemone toxins toward channels Nav1.1–1.7. The presented data demonstrate that the binding sites of these toxins is not restricted to the supposed site 3, between segments S3 and S4 of domain IV. Also, we show that previously assumed critical amino acid positions in this group of peptides may vary and should be carefully considered. By doing this we may avoid misleading interpretations by common generalizations that may arise from site-directed mutagenesis studies. Moreover, subtle variations in primary sequences of toxins may lead to drastic changes in surface charges, such as in the case of δ-AITX-Bcg1a and δ-AITX-Bcg1b, which may contribute to a better understanding of their contact surfaces on the targeted channels.